WO2020008254A1

(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
World Intel 1P v )
O mition % (0 OO
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International Bureau (10) International Publication Number
./
(43) International Publication Date / WO 2020/008254 Al
09 January 2020 (09.01.2020) WIPOIPCT
(51) International Patent Classification: Sylvia; 1021 West Hastings Street, 9th Floor, Vanvouver,
CO7K 16/12 (2006.01) CI2N 1/19 (2006.01) BC V6E 0C3 (CA).
AGIK 39/42 (2006.01) CI2N 1721 (2006.01)
(81) Designated States (unless otherwise indicated, for every
AGIP
CO7K 3112
16/00 (2006.01)
(2006.01) CI2N 1513 (2006.01)
CI2P21/00 2006.01) kind of national protection available): AE, AG, AL, AM,
CO7K 16/08 (2006.01)
i CH0B 40/08 (2006.01)
i AO,
CA, AT, AU,
CH, AZ, BA,
CL, CN, CO, BB,
CR, BG,
CU, BI, BN, DJ,
CZ, DE, BR, DK,
BW, DM,
BY, DO,
BZ,
(21) International Application Number: DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN,
PCT/IB2019/000687 HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP,
KR KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME,
(22) International Filing Date:
MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ,
04 June 2019 (04.06.:2019) OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA.
(25) Filing Language: English SC,SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN,
(26) Publication Language: English
TR, TT. TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
(84) Designated States (unless otherwise indicated, for every
(30) Priority Data: kind of regional protection available): ARIPO (BW, GH,
62/680,736 05 June 2018 (05.06.2018) us GM.KE. LR LS, MW. MZ, NA.RW, SD. SL. ST, §7. TZ.
(71) Applicant: NOVOBIND LIVESTOCK THERA- UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ,
PEUTICS INC. [CA/CAJ; 1021 West Hastings Street, 9th TM), European (AL, AT, BE, BG. CH, CY. CZ, DE, DK,
Floor, Vancouver, Be, VGE OC3 (CA). EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV,
SLOUTET,o Slade,
o, Andrew;
Vo1021 WestBC Hastings
VO oChStreet, 9th TR OAPL(EE B, . G, C CW. G, GN. G0, GW.
(72) Inventors: ABNOUSI, Hamlet; 1021 West Hastings MC, MK, MT.NL, NO. PL. PT. RO. RS, SE. SI. SK. SM.
Floor, Vancouver, BC V6E 0C3 (CA). VAN PETEGEM,

Filip Louis, Arsene; 1021 West Hastings Street, 9th Floor,
Vancouver, BC V6E 0C3 (CA). CHEUNG, Tsz Ying,
(54) Title: ANTIBODIES AGAINST AQUACULTURE DISEASE-CAUSING AGENTS AND USES THEREOF
FIG. 1A
CH2
CH2
w0 20207008254 A1 [l
CH3
CH3
(57) Abstract: Described herein are methods and antibodies useful for reducing, eliminating, or preventing infection with a bacterial
or viral population in an aquatic animal. Also described herein are antigens useful for targeting by heavy chain antibodies and VHH
fragments for reducing a bacterial or viral population in an aquatic animal.
[Continued on next page]

WO 2020/008254 A1 [l
Declarations under Rule 4.17:
— as to applicant's entitlement 1o apply for and be granted a
patent (Rule 4.17(ii))
— as o the applicant's entitlement to claim the priority of the
earlier application (Rule 4.17(iii))
Published:
— with international search report (Art. 21(3))
— before the expiration of the time limit for amending the
claims and to be republished in the event of receipt of
amendments (Rule 48.2(h))
WO 2020/008254 PCT/1B2019/000687
ANTIBODIES AGAINST AQUACULTURE DISEASE-CAUSING AGENTS AND USES

THEREOF
CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 62/680,736, filed
June 5, 2018, which application is incorporated herein by reference. Priority is claimed pursuant
to 35 U.S.C. § 119. The above noted patent application is incorporated by reference as if set
forth fully herein
FIELD OF THE INVENTION
[0002] This invention relates to methods and compositions for the control of microorganisms in
aquaculture and uses thereof.
BACKGROUND OF THE INVENTION
[0003] Losses to the aquaculture industry following contamination of livestock with pathogens
are a global burden. With a growing global population and no significant increase in the amount
of farm land available to agriculture, there is a need to produce larger quantities of food without
using more space. Aquaculture is an especially attractive use of this space because the feed
conversion ratio for aquaculture organisms is roughly 1:1, whereas the ratio for larger farmed
sources of protein is 1:3 or higher . Losses to the global aquaculture industry due to pathogens
is estimated to be around 40%, or $6 billion USD per annum @ Traditional treatment of animals
with antibiotics is a major contributor to the emergence of multi-drug resistant organisms and is
widely recognized as an unsustainable solution to controlling contamination of livestock. There
is a need for the development of pathogen-specific molecules that inhibit infection or association
of the pathogen with the host, without encouraging resistance.
SUMMARY OF THE INVENTION
[0004] With reference to the definitions set out below, described herein are polypeptides
comprising heavy chain variable region fragments (VuHs) whose intended use includes
applications in aquaculture, diagnostics, in vitro assays, feed, therapeutics, substrate
identification, nutritional supplementation, bioscientific and medical research, and companion
diagnostics. Also described herein are polypeptides comprising VyHs that bind to and decrease
the virulence of disease-causing agents in aquaculture. Further to these descriptions, set out
below are the uses of polypeptides that comprise VyHs in methods of reducing transmission and
severity of disease in host animals, including their use as an ingredient in a product. Further
described are the means to produce, characterize, refine and modify VuHs for this purpose
WO 2020/008254 PCT/1B2019/000687
INCORPORATION BY REFERENCE
[0005] All publications, patents, and patent applications mentioned in this specification are
herein incorporated by reference to the same extent as if each individual publication, patent, or
patent application was specifically and individually indicated to be incorporated by reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0006] The novel features of the invention are set forth with particularity in the appended
claims. A better understanding of the features and advantages of the present invention will be
obtained by reference to the following detailed description that sets forth illustrative
embodiments, in which the principles of the invention are utilized, and the accompanying
drawings of which:
FIGS. 1A-1B: Panel A shows a schematic of camelid heavy chain only antibodies and their
relationship to VyH domains. Panel B illustrates the framework regions (FRs) and
complementarity determining regions (CDRs) of the VyH domain
FIGS. 2A-2F: Show phage ELISA binding data for V4H antibodies of this disclosure.
FIG. 3: Shows binding of a selection of recombinantly expressed and purified ViH antibodies
to PirA using a protein pull-down assay.
FIG. 4: Shows the stability of a selection of recombinantly expressed and purified VuH
antibodies to PirA in shrimp midgut extract fluids.
DEFINITIONS
[0007] In describing the present invention, the following terminology is used in accordance with
the definitions below.
[0008] In the following description, certain specific details are set forth in order to provide a
thorough understanding of various embodiments. However, one skilled in the art will understand
that the embodiments provided may be practiced without these details. Unless the context
requires otherwise, throughout the specification and claims which follow, the word “comprise”
and variations thereof;, such as, “comprises” and “comprising” are to be construed in an open,
inclusive sense, that is, as “including, but not limited to.” As used in this specification and the
appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the
content clearly dictates otherwise. It should also be noted that the term “or” is generally
employed in its sense including “and/or” unless the content clearly dictates otherwise. Further,
headings provided herein are for convenience only and do not interpret the scope or meaning of
the claimed embodiments,
WO 2020/008254 PCT/1B2019/000687
1) Host
[0009] As referred to herein, “host”, “host organism”, “recipient animal”, “host animal” and
variations thereof refer to the intended recipient of the product when the product constitutes a
feed. In certain embodiments, the host is a crustacean, a shellfish, a shrimp or a prawn.
2) Shellfish
[0010] As referred to herein, “shellfish” refers to any aquatic exoskeleton-bearing invertebrate

Shellfish can be harvested from the wild or reared. Without limitation, shellfish includes
crustaceans, bivalvia, gastropods, cephalopods, octopus, squid, cuttlefish, clams, oysters,
mussels, scallops, cockles, whelks, winkles, shrimp, prawns, crawfish, crayfish, lobster, crabs,
krill and barnacles.
3) Aquaculture-specific
[0011] As referred to herein, “aquaculture”, “aquatic” and variations thereof refer to the
cultivation or dwelling of organisms, including animals and plants, in water.
4) Pathogens
[0012] As referred to herein, “pathogen”, “pathogenic”, and variations thereof refer to virulent
microorganisms, that can be associated with host organisms, that give rise to a symptom or set
of symptoms in that organism that are not present in uninfected host organisms, including the
reduction in ability to survive, thrive, reproduce. Without limitation, pathogens encompass
parasites, bacteria, viruses, prions, protists, fungi and algae. In certain embodiments, the
pathogen is a bacterium belonging to the Vibrio genus. In certain embodiments, the pathogen is
the White Spot Syndrome Virus.
[0013] “Virulence”, “virulent” and variations thereof refer to a pathogen’s ability to cause
symptoms in a host organism. “Virulence factor” refers to nucleic acids, plasmids, genomic
islands, genes, peptides, proteins, toxins, lipids, macromolecular machineries or complexes
thereof that have a demonstrated or putative role in infection
[0014] “Disease-causing agent” refers to a microorganism, pathogen or virulence factor with a
demonstrated or putative role in infection
5) Bacteria
[0015] As referred to herein, “bacteria”, “bacterial” and variations thereof refer, without
limitation, to Vibrio species, Aeromonas species, Edwarsiella species, Streptococcus species,

Rickettsia species, or any other bacterial species associated with aquatic organisms or host
organisms. In certain embodiments, bacteria may not be virulent in all host organisms it is
associated with
WO 2020/008254 PCT/1B2019/000687
6) Viruses
[0016] As referred to herein, “virus”, “viral” and variations thereof refer, without limitation, to
the White Spot Syndrome Virus, or any other viral species associated with aquatic organisms or
host organisms.
7) Antibodies
[0017] A schematic of camelid heavy chain only antibodies and their relationship to VyH
domains and complementarity determining regions (CDRs) is shown in FIG. 1. (Panel A). A
camelid heavy chain only antibody consists of two heavy chains linked by a disulphide bridge.
Each heavy chain contains two constant immunoglobulin domains (CH2 and CH3) linked
through a hinge region to a variable immunoglobulin domain (ViH). (Panel B) are derived from
single VyH domains. Each VyH domain contains an amino acid sequence of approximately 110-
130 amino acids. The VgH domain consists of the following regions starting at the N-terminus
(N): framework region 1 (FR1), complementarity-determining region 1 (CDR1), framework
region 2 (FR2), complementarity-determining region 2 (CDR2), framework region 3 (FR3),
complementarity-determining region 3 (CDR3), and framework region 4 (FR4). The domain
ends at the C-terminus (C). The complementarity-determining regions are highly variable,
determine antigen binding by the antibody, and are held together in a scaffold by the framework
regions of the VyuH domain. The framework regions consist of more conserved amino acid
sequences; however, some variability exists in these regions.
[0018] As referred to herein “VyH” refers to an antibody or antibody fragment comprising a
single heavy chain variable region which may be derived from natural or synthetic sources.
NBXs referred to herein are an example of a ViH. In a certain aspect a VyH may lack a portion
of a heavy chain constant region (CH2 or CH3), or an entire heavy chain constant region.
[0019] As referred to herein “heavy chain antibody” refers to an antibody that comprises two
heavy chains, and lacking the two light chains normally found in a conventional antibody. The
heavy chain antibody may originate from a species of the Camelidae family or Chondrichthyes
class. Heavy chain antibodies retain specific binding to an antigen in the absence of any light
chain
[0020] As referred to herein “specific binding”, “specifically binds” or variations thereof refer to
binding that occurs between an antibody and its target molecule that is mediated by at least one
complementarity determining region (CDR) of the antibody’s variable region. Binding that is
between the constant region and another molecule, such as Protein A or G, for example, does not
constitute specific binding.

[0021] As referred to herein “antibody fragment” refers to any portion of a conventional or
heavy chain antibody that retains a capacity to specifically bind a target antigen and may include
WO 2020/008254 PCT/1B2019/000687
a single chain antibody, a variable region fragment of a heavy chain antibody, a nanobody, a
polypeptide or an immunoglobulin new antigen receptor (IgNAR)
[0022] As referred to herein an “antibody originates from a species” when any of the CDR
regions of the antibody were raised in an animal of said species. Antibodies that are raised in a
certain species and then optimized by an in vitro method (e.g., phage display) are considered to
have originated from that species.
[0023] As referred to herein “conventional antibody” refers to any full-sized immunoglobulin
that comprises two heavy chain molecules and two light chain molecules joined together by a
disulfide bond. In certain embodiments, the antibodies, compositions, feeds, products, and
methods described herein do not utilize conventional antibodies

8) Production System
[0024] As referred to herein, “production system” and variations thereof refer to any system that
can be used to produce any physical embodiment of the invention or modified forms of the
invention. Without limitation, this includes but is not limited to biological production by any of
the following: bacteria, yeast, algae, arthropods, arthropod cells, plants, mammalian cells.
Without limitation, biological production can give rise to antibodies that can be intracellular,
periplasmic, membrane-associated, secreted, or phage-associated. Without limitation,
“production system” and variations thereof also include, without limitation, any synthetic
production system. This includes, without limitation, de novo protein synthesis, protein
synthesis in the presence of cell extracts, protein synthesis in the presence of purified enzymes,
and any other alternative protein synthesis system.
9) Product
[0025] As referred to herein, “product” refers to any physical embodiment of the invention or
modified forms of the invention, wherein the binding of the Vy4H to any molecule, including
itself, defines its use. Without limitation, this includes a feed, a feed additive, a nutritional
supplement, a premix, a medicine, a therapeutic, a drug, a diagnostic tool, a component or

entirety of an in vitro assay, a component or the entirety of a diagnostic assay (including
companion diagnostic assays).
10) Feed Product
[0026] As referred to herein, “feed product” refers to any physical embodiment of the invention
or modified forms of the invention, wherein the binding of the VyH to any molecule, including
itself, defines its intended use as a product that is taken up by a host organism. Without
limitation, this includes a feed, a pellet, a feed additive, a nutritional supplement, a premix, a
medicine, a therapeutic or a drug.
WO 2020/008254 PCT/1B2019/000687
DETAILED DESCRIPTION OF THE INVENTION

[0027] Descriptions of the invention provided are to be interpreted in conjunction with the
definitions and caveats provided herein.
[0028] Some farmed aquatic organisms, such as some crustaceans, lack a true adaptive immune
response. Additionally, the administration of therapeutics by injection for small and intensely
reared organisms is cumbersome. For these reasons, vaccine-based approaches to protecting
farmed aquaculture organisms from pathogenic infection is ineffective. Secondly, the use of
antibiotics as growth promoters in animal feed has already been banned in Europe (effective
from 2006) in an effort to phase out antibiotics for non-medicinal purposes and limit
antimicrobial resistance. Indeed, many bacterial pathogens of aquatic organisms already harbor
resistance to common antibiotics. This underpins the need for the development of non-antibiotic
products to administer to aquatic organisms to prevent infection and promote growth.
[0029] Significant pathogens affecting farmed aquatic organisms include bacteria, such as
members of the Vibrio genus, among others, as well as viruses such as White Spot Syndrome
Virus (WSSV). Losses due to Vibrio parahaemolyticus, for example, first emerged in 2009 and
have been prevalent ever since ™. It was not until 2013 that V. parahaemolyticus was shown to
be the causative agent of Acute Hepatopancreatic Necrosis Disease (AHPND): a subtype of
Early Mortality Syndrome (EMS) that contributes approximately $1 billion USD loss to the
shrimp farming industry per annum “ . In 2015 it was demonstrated that the presence of the
PVA-1 plasmid and the toxins encoded (PirA and PirB) are directly responsible for AHPND
Once infected, organisms are up to 100% moribund within 3 days. V. parahaemolyticus is also a
prevalent human pathogen, responsible for gastrointestinal infection and septicemia after
exposure to contaminated fish or fisheries®. In addition to PirA and PirB, V. parahaemolyticus
produces several proteinaceous factors that have been demonstrated to facilitate host infection
and can be targeted to curb virulence.
[0030] WSSV infection is a longer-standing problem; having been identified in 1992 7 there is
still no effective means of controlling viral spread or infection in aquatic organisms. Cumulative
losses to the aquaculture industry as a consequence of WSSV are estimated at $15 billion USD
® Infected organisms are moribund within 3-5 days. The surface of the viral envelope is well
characterized and can be targeted to prevent infection.
[0031] Other disease-causing agents affecting farmed aquaculture organisms include bacteria
(such as Yersinia spp., Edwarsiella spp., Aeromonas spp., Streptococcus spp. and Rickettsia
spp.), viruses (such as White Spot Syndrome Virus (WSSV), Yellowhead virus, tilapia
iridovirus, epizootic hematopoietic necrosis virus (EHNV), infectious hematopoietic necrosis
virus (IHNV), infectious salmon anemia virus (ISAV), infectious pancreatic necrosis virus
WO 2020/008254 PCT/1B2019/000687
(IPNV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), taura syndrome
virus (TSV) and white spot bacilloform virus (WSBV), hepatopancreatic parvo-like virus
(HPV), reo-like virus, monodon baculovirus (MBV), baculoviral midgut GI and necrosis virus
(BMN)), algae, prions, protists, parasites, fungi, peptides, proteins and nucleic acids. To our
knowledge, an effective, non-vaccine-based treatment against any of these disease-causing
agents has yet to be developed for commercial use.
[0032] Existing methods fail to acknowledge the limited immune development of aquatic
organisms affected by the pathogens listed above, and as such rely on the host organism to
generate protection against disease-causing agents. This approach is limited by the inadequacies
of the host organism’s immune system and therefore does not provide an effective means of
protection. This problem is circumvented by introducing exogenous peptides into the host that
neutralize the virulence and spread of the disease-causing agent without eliciting the host
immune response. Moreover, the methods described herein provide scope for the adaptation and
refinement of neutralizing peptides, which provides synthetic functionality beyond what the host
is naturally able to produce.
[0033] Antibody heavy chain variable region fragments (VyHs) are small (12-15 kDa) proteins
that comprise specific binding regions to antigens. When introduced into an animal, ViHs bind
and neutralize the effect of disease-causing agents i situ. Owing to their smaller mass, they are
less susceptible than conventional antibodies, such as previously documented IgYs, to cleavage
by enzymes found in host organisms, more resilient to temperature and pH changes, more
soluble, have low systemic absorption and are easier to recombinantly produce on a large scale,
making them more suitable for use in animal therapeutics than conventional antibodies
Antibodies for preventing or reducing virulence (summary)
[0034] In one aspect, the present invention provides a polypeptide or pluralities thereof
comprising a VyH or VyHs that bind disease-causing agents to reduce the severity and
transmission of disease between and across species. In certain embodiments, the VyH is
supplied to host animals. In certain embodiments, the VyH is an ingredient of a product.
Binding to reduce virulence
[0035] In another aspect, the present invention provides a polypeptide or pluralities thereof
comprising a VygH or VyHs that bind disease-causing agents, and in doing so, reduce the ability
of the disease-causing agent to exert a pathological function or contribute to a disease
phenotype. In certain embodiments, binding of the V4H(s) to the disease-causing agent reduces
the rate of replication of the disease-causing agent. In certain embodiments, binding of the
ViH(s) to the disease-causing agent reduces the ability of the disease-causing agent to bind to its
cognate receptor. In certain embodiments, binding of the Vy4H(s) to the disease-causing agent
WO 2020/008254 PCT/IB2019/000687
8
reduces the ability of the disease-causing agent to interact with another molecule or molecules
In certain embodiments, binding of the VyH(s) to the disease-causing agent reduces the mobility
or motility of the disease-causing agent. In certain embodiments, binding of the VyzH(s) to the
disease-causing agent reduces the ability of the disease-causing agent to reach the site of
infection. In certain embodiments, binding of the VyH(s) to the disease-causing agent reduces
the ability of the disease-causing agent to cause cell death.
Antibodies derived from llamas
[0036] In a further aspect, the present invention provides a method for the inoculation of
Camelid or other species with recombinant virulence factors, the retrieval of mRNA encoding
VuH domains from lymphocytes of the inoculated organism, the reverse transcription of mRNA
encoding VyH domains to produce cDNA, the cloning of cDNA into a suitable vector and the
recombinant expression of the VyH from the vector. In certain embodiments, the camelid can be
a dromedary, camel, llama, alpaca, vicuna or guanaco, without limitation. In certain
embodiments, the inoculated species can be, without limitation, any organism that can produce
single domain antibodies, including cartilaginous fish, such as a member of the Chondrichthyes
class of organisms, which includes for example sharks, rays, skates and sawfish. In certain
embodiments, the heavy chain antibody comprises a sequence set forth in Table 1. In certain
embodiments, the heavy chain antibody comprises an amino acid sequence with at least 80%,
90%, 95%, 97%, 99%, or 100% identity to any sequence disclosed in Table 1. In certain
embodiments, the heavy chain antibody possesses a CDRI1 set forth in Table 2. In certain
embodiments, the heavy chain antibody possesses a CDR2 set forth in Table 2. In certain
embodiments, the heavy chain antibody possesses a CDR3 set forth in Table 2
Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

SEQ
D NBX Amino acid sequence Antigen
1 NBX0401 QVQLQQSGGGLVRAGGSLRLSCETSGRTFSSYTMG PirA
WFRQAPGKEREFVGTIDWW SSSSSYADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCAASGKYGLA
YSRRDYAYWGQGTQVTVSS
2 NBX0402 QVQLQQSGGSLVQAGGSLRLSCAASGLPFINYAMG PirA
WFRQAPGKDREIVAAIDWNGDSTYYAVSVKGRFTI
SRDNAKNTVTLQMNSLKPEDTAIYYCASHYQPYIR
VSATRRFEADYWGQGTQVTVSS
3 NBX0403 | QVQLQQSGGSLVQAGGSLRLSCAASGLPFINYAMG | PirA
WFRQAPGKDREIVAAIDWNGDSTYYAVSVKGRFTI
SRDNAKNTVTLQMNSLKPEDTAIYYCAADYQPYIR
VSATRRFEADYWGQGTQVTVSS
4 NBX0404 QVQLQESGGGLVQAGDSLRLSCATSGRTFSRYTMG PirB
WFRQTPGKEREFVAAISWSGTYYTDSVKGRFTISV
DNAKNTVYLQMNSLKPEDTAVYYCASGSRRLYYS
WO 2020/008254 PCT/1B2019/000687

SEQ
NBX Amino acid sequence Antigen
SDIDYWGQGTQVTVSS
NBX0405 QVQLQESGGGLVQAGDSLRLSCATSGRTFSRYTMG PirB
WFRQTPGKEREFVAAISWSGTYYTDSVKGRFTISRD
NAKNTVYLQMNSLKPEDTAVYYCVVGSRRLYYSS
DINYWGQGTQVTVSS
NBX0406 QVQLQESGGGLVQAGESLRLSCAASGFTFSTYTWD VP24
WYRQAPGKQREMVARISRDGITNY ADSVKGRFTIS
RDNAKNTVDLQMNSLKPEDTAVY YCAVVKEDNR
YWCHADRNLYRNWGQGTQITVSS
29 NBX0601 QVQLQQSGGGLVQPGGSLRLSCAGSRFTFSTYPMS PirA
WVRQAPGKGVEWVSSISVGGGIKNYADSVKGRFTI
SRDNAKNTMYLQMNGLKPEDTAVYYCAKGGKTS
YTREWGQGTQVTVSS
NBX0602 QVQLQESGGGLVQAGDSLRLSCAASGRTFSRYAM PirA
GWFRQAPGKEREFVAAVDWSGGSTAYADSVKGRF
TISRDNAKNTVYLQMNSLKPDDTAVYYCAARARD
VYGRAWYVEDSSTYDYWGQGTQVTVSS
NBX0603 QVQLQESGGGLVQAGGSLRLSCAASGSMFSINAMG PirA
WYRQAPGNEREW VATISRGGITYYDDSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYFCNAENRRLGD
DFWGQGTQVTVSS
32 NBX0604 QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYTMG PirA
WFRQVPGKEREFVAAIRWSGGSRIYADSVKGRFTIS
RDNTKNVVYLQMTSLKPEDTAVYYCAADRDGYSS
YAHQYDYWGQGTQVTVSS
33 NBXO0605 QVQLQESGGGLVQPGGALRLSCAASGSFFSIYAMA PirA
WYRQAPGKQRELVAGITSGSETNYADSVK GRFTIS
RDNAKNTVYLQMNSLKLEDTAVYYCNRWAPLTRL
DYWGQGTQVTVSS
34 NBX0606 QVQLQESGGGLVQAGDSLRLSCAASGRTSSSFAMG PirA
WFRQAPGKEREFVGGITRTGGRTY YVDSVKGRFTI
SRDNAKNTMSLQMNSLKPEDTAVYYCAARWATA
TSNSIRVYYNEGQYDYWGQGTQVTVSS
35 NBX0607 QVQLQESGGGLVQAGGSLRLSCAASGGTFSRLTMG PirA
WFRQAPGEEREFVAAVSWVAETTDYADSVKGRFSI
SRDNSKNTVYLQMNSLKPVDTAVYYCAAGPRDMI
RSRNIRSYDSWGQGTQVTVSS
36 NBX0608 QVQLQESGGGLVQAGGSLRLSCAASGSMFNINAM PirA
GWYRQAPGKQREWVATISSGGITY YDDSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAVYFCNAENRRLG
DDYWGQGTQVTVSS
37 NBX0609 QVQLQESGGGLVQAGGSLRLSCAASGSIFSGNVMG PirA
WYRQVPGKLREWVATISSGGITYYDDSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYFCNLENRRLGD
DYWGQGTQVTVSS
38 NBX0610 QVQLQESGGGLVQAGGSLRLSCAASGSIFSRDAMG PirA
WYRQAPGNLREWVATISSGGITYYDDSVKGREFTIS
RDNAKNTVYLQMNSVKPEDTAVYFCNAENRRLGD
WO 2020/008254 PCT/1B2019/000687
10

DYWGQGTQVTVSS
NBX0611 QVQLQQSGGGLVQAGGSLRLSCAASGGTFSRLTM PirA
GWFRQAPGEEREFVAAVSWVAETTDYADSVKGRF
SISRDNSKNTVYLQMNSLKPVDTAVYYCAAGPRD
MIRSRNIKSYDSWGQGTQVTVSS
40 NBX0612 QVQLQESGGGLVQAGGSLRLSCVASGTIFSINKMG PirA
WYRQAPEKERELVAVARSGGIINYADSVKGRFTISR
DDAKNTVYLQMNSLRPDDTAVYFCNALIHTRYDR
VIGYWGQGTQVTVSS
41 NBX0613 QVQLQESGGGLVQAGGSLRLSCAASGSIFSINAMG PirA
WYRQAPGKLREWVATISSGGITYYDDSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYFCNAENPRLGD
DYWGQGTQVTVSS
2 NBX0614 QVQLQQSGGGLVQAGGSLRLSCAASGSSFRSNAIG PirB
WYRQFPGKSRELIAVITRSGSTQYADSVKGRFTASR
DNAKNMIYLQMNNLKLEDTAVYYCHDETMKLISV
KNDYWGQGSQVTVSS
43 NBX0615 QVQLQESGGGLVQAGGSLRLSCAASGSMFSRNAM PirA
SRDNAKNTVYLQMNSLKPEDTAVYFCNAENRRLG
DDYWGQGTQVTVSS
44 NBX0616 QVQLQESGGGLVQAGGSLRLSCATSGLTFSSYAMG PirA
WFRQAPGKEREFVATISWSGKSTRYSDSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCAADYQRLGL
LRVGVAEYDYWGQGIQVTVSS
45 NBX0617 QVQLQQSGGGLVQAGGSLRLSCQASGRSGSTSFMG PirA
WFRQAPGKEREFVAAIRWSSGMTY YADSVKGRFTI
SRDNAKNTVDLQMNSLKPEDTAVYYCAADNYPLH
IGHQDHEVDYWGQGTQVTVSS
6 NBX0618 QVQLQESGGGLVQPGGSLRLSCAASGSIFSFNAMG PirA
WYRQAPGKQRELVAAITKGGSTSYADSVKGRFTIS
VDNAKNTVYLQMNSLTPEDTAVYYCNVKTLRSAL
FPGYEYWGQGTQVTVSS
47 NBX0619 QVQLQQSGGGLVQAGGSLRLFCAASGGTFSRLTM PirA
GWFRQAPGEEREFVAAVSWVAETTDYADSVKGRF
SISRDNSKNTVYLQMNNLKPVDTAVYYCAAGPRD
MIRSRNIRSYDSWGQGTQVTVSS
43 NBX0620 QVQLQESGGGLVQAGGSLRLSCAASGGTFSRLTLG PirA
WFRQAPGEEREFVAAVSWVAEMTDYADSVKGRFS
ISRDNSKNTVYLQMNSLKPVDTAVYYCAAGPRDM
VRSRNIRSYDSWGQGTQVTVSS
49 NBX0621 QVQLQESGGGLVQAGGSLRLSCAASGRTFSTYSMG PirA
WFRQVPGKEREFVAAIRWSGGSRTYADSVKGRFTI
SRDNTKNVVYLQMTSLKPEDTAVYYCAADRDGYS
SYAHQYDYWGQGTQVTVSS
50 NBX0622 QVQLQESGGGLVQAGGSLRLSCAASGRLFNINAMG PirA
WYRQAPGKQREW VATISSGGITY YDDSVKGRFTIS
RDNAKNTVYLQMDSLKPGDTAVYFCNAENRGLGD
WO 2020/008254 PCT/1B2019/000687
11

SEQ
DYWGQGTQVTVSS
51 NBX0623 QVQLQESGGGWVQTGGSLRLSCAASGRTLSNYAM PirA
GWFRQAPGKEREFVAAISRSGMSTDAPNSVKGRFT
VSRDNAKNTMYLHLNSLKPEDTAVYYCAARGGLP
NPSRTYGFEEQYDYWGQGTQVTVSS
52 NBX0624 QVQLQESGGGLVQAGGSLRLSCAASGRTVSSLPMG PirA
WFRQAPGKEREFVAALNWSGTSTYYEDSVKGRFTI
SRDNAKNTLYLQMNNLKPEDTAVYYCAAKGAIYY
SYSPRNNNNYDVWGQGTQVTVSS
53 NBX0625 QVQLQESGGGLVQAGGSLRLSCAASGSMFNINAM PirA
SRDNAKNTVHLQMNSLKPEDTAVYFCNAENRLGD
DYWGQGTQVTVSS
54 NBX0626 QVQLQQSGGGLVQAGGSLRLACAVSETTLATNAM PirB
AWYRQAQGKRREW VATISSVSSGGITNYSGSVKGR
FTISRDNAKNTVFLQMNSLQPEDTAVYYCNGVRRG
RSYWGQGTQVTVSS
55 NBX0627 QVQLQQSGGGLVQAGGSLRLSCAASGSSFRSNAIG PirB
WYRQSPGKSRELIAVITRSGSTQYADSVKGRFTASR
DNAKNMIYLQMNSLKPEDTAVYYCHDETMKLITG
KNDYWGQGTQVTVSS
56 NBX0628 QVQLQQSGGGLVQVGGSLRLSCAASGRTFSSYAM PirA
GWFRQVPGKEREFVAAIKWSGGSRTYADSVAGRF
TISRDNTKNVVYLQMTSLKPEDTAVYYCAADRDG
YSRYAHQYDYWGQGTQVTVSS
57 NBX0629 QVQLQQSGGGLVQAGGSLRLSCAASGGTFSRLTIG PirA
WFRQAPGEERVFVAAVSWVAETTDYADSVKGRFSI
RSRNIRSYDSWGQGTQVTVSS
58 NBX0630 QVQLQQSGGGLVPGGGSLRLSCAASGVTFSDYPMA PirA
WYRTAPGKQRELVASISAGGGLIKY VDSVKGRFTIS
RDNAKNTLYLQMNSLKPEDTGVYLCNLKTSYFWP
WGQGTQVTVSS
59 NBX0631 QVQLQESGGGLVQAGDSLRLSCKASGGTFSRLTIA PirA
WFRQAPGKEREFVTAVSWVAQTTDYADSVKGRFSI
SRDNSKNTVYLQMNSLKPLDTAVYYCAAGPQDMI
RSRNIRSYISWGQGTQVTVSS
60 NBX0632 QVQLQQSGGGLVQAGGSLRLSCAASGGTFSRLTM PirA
GWFRQAPGEEREFVAAVSWVAGTTDYADSVKGRF
TISRDNSKNTVHLQMNSLKPVDTAVYYCAAGPRD
MIRSRNIRSYDSWGQGTQVTVSS
61 NBX0633 QVQLQESGGGLVQPGGSLRLSCTASGTIFRSKSMA PirA
WYRQAPGQGRETVAHISGLGHTNY VESVKGRFTVS
RDDAKNAVYLQMSSLKPEDTAVYYCNTFTAAFSW
GQGTQVTVSS
62 NBX0634 QVQLQQSGGGLVQAGGSLRLSCAASGGSFSRLTLG PirA
WO 2020/008254 PCT/1B2019/000687
12

SEQ
RSRNIRSYASWGQGTQVTVSS
63 NBX0635 QVQLQESGGGLVQAGGSLRLSCATSGLTFSNYAMG PirA
WFRQAAGKEREFVATISWSGKSTRYADSVKGRFTI
SRDNAKNTVDLRMNSLKPEDTAVYYCAAEYQRLG
LLRDGVADYSYWGQGTQVTVSS
64 NBX0636 QVQLQESGGGLVQAGGSLRLSCAASGGTFSRLTMG PirA
RSRNIRSYVSWGQGTQVTVSS
65 NBX0637 QVQLQESGGGLVQAGNSLKLSCVASGRTFSSYPMG PirA
WYRTAPGKQRELVASISAGGGLIKY VDSVKGRFTIS
RDNAKNTLYLQMNSLKPEDTGVYLCNLKTSYFWP
WGQGTQVTVSS
66 NBX0638 QVQLQESGGGLVQAGGSLRLSCAASGSIFSGNVMG PirB
WYRQVPGKQRDLVATITGGGITRY ADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCNYRRIMQQY
WGKGTLVTVSS
67 NBX0639 QVQLQESGGGLVQAGGSLRLSCAASGIVFSSIVMA PirB
WYRQAPGKQRELVASITNGGLVNSGDSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCNARRIMTSY
WGQGTQVTVSS
68 NBX0640 QVQLQESGGGLVQPGGSLRLSCAASGSIFSGNVMG PirB
WYRQVPGKQRDLVATITGGGITRYADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCHYRRIMQQY
WGQGTQVTVSS
69 NBX0641 QVQLQESGGGLVQAGGSLRLSCAASGSISNSYVMG PirB
WYRQAPGKQRELVATITSGGLTNYAQSLKGRFTIS
RDNAKNTVYLQMTSLEPEDTAVYYCNARVIFTTY
WGQGTQVTVST
70 NBX0642 QVQLQQSGGGLVQAGGSLRLSCVASTVIFSRYAM PirB
GWFRQAPGKEREVVAGISGSGHRTYYGDFVKGRFT
ISRDNAKKTVYLQMNNLKPEDTAVYYCAGDLVAK
FDSAYRVSYDSWGQGTQVTVSS
71 NBX0643 QVQLQESGGGLVQAGGSLRLSCEASGSIFSGNVMG PirB
WYRQVPGKQRDLVATMTGGGVTRYADSVKARFTI
SRDNAKNTVYLQMNSLKPEDTGVYYCHYRRIMQQ
YWGQGTQVTVSS
72 NBXO0644 QVQLQESGGGLVQAGGSLRLSCAASGSIFSGNVMG PirB
RDNAKNKVYLQMNGLKPEDTAVYFCFYRRIMQQY
WGQGTQVTVSS
WYRQVPGKQRDLVATITGGGITHYADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCLYRRIMQQY
WGQGTQVTVSS
WYRQVPGNQRELVATITGGGVTRYADSVKARFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCHYRRIMQQS
WO 2020/008254 PCT/1B2019/000687
13

SEQ
WGQGTQVTVSS
75 NBX0647 QVQLQESGGGSVPPGGSLRLSCAASGSIFSGNVMA PirB
WYRQVPGKQRDLVASMTGGGVTRYADSVIARFTIS
RDNVKNTVYLQMNSLKPEDTAVYYCHYRRIMQQY
WGQGTQVTVSS
76 NBX0648 QVQLQESGGGLVQAGGSLRLSCEASGIIFSSNVMG PirB
WYRQAPGKQRELVASRTSGGLTNYADSAKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCNARRLFTNY
WGQGTQVTVSS
77 NBX0649 QVQLQQSGGGLVQPGGSLTLSCAASGSIASGNVLG PirB
WYRQAPGKQRELVATITSGGLTHYKDSVKGRFTIS
RDNAKNMVFLQMNSLKPEDTAVYYCNYRRLATG
YWGQGTQVTVSS
78 NBX0650 QVQLQESGGGLVQAGGSLRLSCEASGSIFSGNVLG PirB
RDNAKNTVYLQMNALKPEDTAVYYCHYRRIMQQ
YWGQGTQVTVSS
79 NBX0722 QVQLQESGGGLVQAGGSLRLSCRASGRTFSSYPMG VP28
WFRQAPGKEREQIAGISRSGDPGKYAASVSGRFTIS
RDNAKNTLYLQMNSLKPEDTAVYYCAARQIYSNN
YSYWGQGTQVTVSS
80 NBX0723 QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYPMG VP28
WFRQAPGSEREQIAGISRSGVPGKYADSVSGRFTIS
RDNAKNTLYLQMNSLKPEDTAVYYCAARSIYSNN
YSYWGQGTQVTVSS
81 NBX0724 QVQLQQSGGGLVQAGGSLRLSCAASGRTFSSYPMG VP28
WFRQAPGSEREQIAGISRSGVSGKYADSVSGRFTIS
RDNAKNTLYLQMNSLKPEDTAVYYCAARSIYSNN
YSYWGQGTQVTVSS
82 NBX0725 QVQLQQSGGGLVQAGGSLRLSCTASGRTFSSYPMG VP28
WFRQAPGKEREQIAGISRSGNPGKYADSVSGRFTIS
YSYWGQGTQVTVSS
83 NBX0730 QVQLQESGGGLVQTGDSLRLACAASGRTFSSYPMA VP28
WYRQAPGQEREFVAGINRNGNIPVY ADSVKGRFTI
SRDNAKNTVYLQMNNLKPEDTAVYYCAARTIYDS
HYTSWGQGTQVTVSS
34 NBX0737 QVQLQQSGPGLVKPSETLSLTCTVSDGAITGSYYV VP24
WSWIRQPPGKGLEWMGVITYDGSTYYNPSLESRTSI
SRDTSKNQFSLQLDSVTREDTAVYYCARAGEEYICS
GYGCHGSLGLDYWGKGTLVTVSS
85 NBXO0738 QVQLQESGGGLVQPGGSLGLSCAASGFTFGSYAMS VP24
WVRQAPGKGPEW VSGINSGDGRITYADSVKGRFTI
SRDNTKNTLYLQMNSLKPEDTAVYYCATGIRTPIIW
GQGTQVTVSS
86 NBX0739 QVQLQESGGGLVQAGGSLRLSCAASGSIFTSDVGW VP24
NRQAPGSVREVVARMTSAGTTIYGDDVMGREFTISR
DNAKSTVYLQMNSLLPEDTGVYYCGVGRFWGQGT
WO 2020/008254 PCT/1B2019/000687
14

SEQ
QVTVSS
87 NBXO0745 QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYPMG VP28
WFRQAPGKEREQIAGISRSGDPGKYAASVSGRFTIS
RDNAKNTLYLQMNSLKPEDTAVYYCAAREIYSNN
YSYWGQGTQVTVSS
88 NBX0746 QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYAMG VP28
WFRRAPGKEREQIAGISRSGNPGKYADSVSGRFTIS
YSYWGQGTQVTVSS
89 NBX0813 QVQLQESGGGLVQAGGSLRLSCVVSGMLFSIRNMR PirA
WYRQAPGKQRELVAQIGSSGNTDY VESVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYFCNALNYWGK
GTLVTVSS
90 NBX0814 QVQLQQSGGDLVQAGGSLRLSCAASMRTFNSRTIG PirA
WFRQAPGKGRELAAAIAWTGGNTYYADSVKGRFT
ISRDNAKNTVYLQMNSLKPEDTAVYYCAAQTRPY
DLPSIRPDDYASRGQGTQVTVSS
91 NBX0815 QVQLQESGGGLVQAGGSLRLSCAASGRTFSSYAMG PirA
WFRQSPGKDREFVAAVSWSGGSTYYADSLKGRFTI
SRDNAKNTVYLQMNSLKPEDTADYYCAAQRVMD
YYRPRTESAYAYWGQGTRVTVSS
92 NBX0816 QVQLQESGGGLVQAGGSLRLSCAASEYIFSNFGMG PirA
WFRQAPGKEREFVGAISRSGSRMSYADSVKGRFIIS
RDNTKNTVYLQMNSLKPEDTAVYYCAAVYGQYSY
HYSSDSKQYSYWGQGTQVTVSS
93 NBX0817 QVQLQESGGGLVQAGGSLRLSCGASGGTNSNYAM PirA
GWFRQPPGKKREFVAALSWSGYNTHYADSVKGRF
TISRPSARTVDLQMNNVKPEDTAVYYCAARLSGRT
AGSRTYYAEGQYDYRGQGTQVTVSS
94 NBX0818 QVQLQQSGGGLVQAGGSLRLSCAASGRTSSSSYLG PirA
WFRQAPGKEREFVASIRWSDGSTYYRDSVEGRFTIS
RDNAKNTVYLRMNSLKPEDTAVYYCAAATTDWG
PRGPYNYWGQGTLVTVSS
95 NBX0819 QVQLQQSGGREVRPGDSLRLSCRASGRTSGAWNM PirA
AWFRQAPGKDREFVAAISGSGRTTEYADSAKGRFT
ISRDMAKNTVYLQMNSLKPEDTAVYNCAASTFDW
GPRGPYRLWGQGTQVTVSS
96 NBX0820 QVQLQESGGGLVQTGGSLRLSCAASGRTFSNYVIG PirA
WFRQAPGKEREFVAAVGRGINSAYHATHYSESVK
DRFTTSRDNAKNTGFLQMNSLKTEDTAVYYCAVTS
RWGQFDRTDFNSWGQGTQVTVSS
97 NBX0821 QVQLQESGGGLVQAGGSLRLSCVVSGMLFSIRNMR PirA
WYRQAPGKQRELVAQIGTSGATDY VGSVEGRFTIS
RDNPKNTVYLQMNSLKPEDTAVYFCNALNYWGEG
TLVTVSS
98 NBX0822 QVQLQESGGGLVRPGDSLTLSCTYSGQTFTNSGMA PirA
WFRQRPGKEREFVAAVSRSGLGRRYADSVRGRFTI
TRDNGKNTANLQMDSLKPEDTAVYSCAATTLDWG
WO 2020/008254 PCT/1B2019/000687
15

SEQ
PRGPYRYWGQGTQVTVSS
99 NBX0823 QVQLQQSGGGLVQTGGSLRLSCAASGSIFSIDFMG PirA
WYRQAPGNPREFVARIRGGNTYYADSVKGRFNISR
DNAENTVYMQMNSLKSEDTAVYYCNAQITMRGGT
WSTSEYWGQGTQVNVSS
100 NBX0824 QVQLQESGGGLVQAGGSLRLTCAASGRTLSSYLSS PirA
YAMGWFRQAPGKERESVATITWNGDRTLYADAVK
GRFTISRDNAKNTVYLQMNSVIPEDTAVYYCAADT
VGRWRSTLSVRDEYDYWGQGTQVTVAS
101 NBX0825 QVQLQESGGGVVETGGSLSVSCVASGRTFSAYTMA PirA
WFRQSPGKEREFVASMSRGSAAYYTDSVRGRFAIS
RVGDKNTVHLQMRDLKPEDTAVYYCAGGSPGSSQ
IATPEAYTYWGQGTQVTVSA
102 NBX0826 QVQLQESGGGLVQAGGSLRLSCAASGSISSSHVMG PirB
WYRQAPGKQRELVATITSGGSTIYADSVKGRFTVS
RDNAKNTVYLQMNSLKSEDTAVYYCHARRLWNT
YWGQGTQVTVSS
NBX0827 QVQLQESGGGLVQAGGSLRLSCAASGSISSSFVMG PirB
YWGQGTQVTVSS
104 NBX0828 QVQLQESGGGAVQAGGSLRLSCAGPRSIFSGNAMA PirB
WYRQVPGKQRETVATVNTGGLTWYGDFVKGRFTI
SRDDAKNTLLLQMDSLKPEDTAVYYCNAVLVRAR
GMWGQGTQVTVSS
105 NBX0829 QVQLQESGGGSVQPGGSLRLSCSASGDRLSSYVMG PirB
WYRQAPGKQRELVATVTSGGRTNYADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCNARILFTNY
WGQGTQVTVSS
106 NBX0830 QVQLQESGGGLVQPGGSLQLSCVASGSVLSRYVM PirB
GWYRQAPGKQRELVATITSGGITRY ADSMKGRFTI
SRDNAKNTVHLQMSSLKPEDTAVYYCNARALWNT
YWGQGTQVTVSS
107 NBX0831 QVQLQESGGGLVQAGGSLRLSCSASGDAFSRYVM PirB
GWYRQAPGKQRELVATITSGGSTIYADSVKGRFTV
SRDNAKNTVYLQMNSLKSEDTAVYYCHARRLWNT
YWGQGTQVTVSS
108 NBX0832 QVQLQQSGGGLVQAGGSLRLSCAASGSISSSYVMG PirB
YWGQGTQVTVSS
109 NBX0833 QVQLQQSGGGLVQAGGSLRLSCSASGSISSSHVMG PirB
RDNAKNTVYLQMNSLKSEDTAVYYCHARRLWDT
YWGQGTQVTVSS
110 NBX0834 QVQLQESGGGSVQAGGSLRLSCAASESMFRDHNM PirB
GWYRQAPGKQRELVATISRGGLINYGDSVRGRFTIS
RDNAKNTIYLQMNSLKVEDTAVYYCNARRLLTTV
WO 2020/008254 PCT/1B2019/000687
16

SEQ
WGQGTQVTVSP
111 NBX0835 QVQLQESGGGLVQPGGSLRLSCSASGNRFSSSYVM PirB
GWYRQAPGKQRELVATVTSGGLTHFKDSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAVYYCNARILLTN
YWGQGTQVTISS
112 NBX0836 QVQLQQSGGGLVQPGGSLRLSCSASGIRLGSY VMG PirB
YYRQAPGKQRELVATVTSGGTTNRADSVKGRFTIS
RDNAKNAVYLQMNSLKPEDTAVYYCNARILFTNY
WGQGTQVTVSS
NBX0837 QVQLQESGGGLVQPGGSLRLSCAASGIVFANHVMG PirB
WYRQAPGKQRELVASITNGGLINSVDSVAGRFTISR
DNAKNTVYLQMNNLKPEDTAVYYCNARRLYQQY
WGQGTQVTVSS
114 NBX0838 QVQLQESGGGLVQAGGSLRLSCRVSGRTVGSYAM PirB
GWFRLQPGKERQFVAAIGWSGASTLYAESVKGRFT
ISRDNAKNTVYLQMNSLKPEDTAVYYCAQRPSSRY
ASRYLGDYAYWGQGTQVTVSS
115 NBX0839 QVQLQESGGGLVQAGGSLRLSCAASGSIGSDYVLG PirB
WYRQAPGKQRELVATITSGGLTHYGDSVKGRFTIS
RDNAKNTVYVQMNSLKFEDTAIYYCNARRLFRNY
WGQGTQVTVSS
116 NBX0840 QVQLQQSGGGLVQAGGSLRLSCAASGSIRSSNVMG PirB
WYRQTPGKQRELVATMTAGGLTNYADSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAVYYCHYRRIFNV
YWGQGTQVTVSS
117 NBX0841 QVQLQESGGGLVQAGGSLRLSCAASARTFIYKMG PirB
WFRQAPGKERDFVASIMWSVGNNYYYTDSAKGRF
TISRDIAKNTMYLQMDSLEPEDTGEYYCAAATTST
QWRYWGQGTQVTVSS
118 NBX0842 QVQLQESGGGWVQPGGSLRLSCAASGSIDNGYVM PirB
GWYRQAPGKQRELVATITSGTNTHYADSVKGRFTI
SRDNAKTTVYLQMNSLKPEDTAVYYCLARRLFTM
YWGQGTQVTVSS
GYYRQAPGKQRELVATVTTGGLTNYADSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAVYYCNARILLTN
YWGQGTQVTVSS
120 NBX0844 QVQLQESGGGLAQTGDSLRLSCAASGRMFSGFVM PirB
GWYRQNPGKQRELVATITNGGLTHYGDSVKGRFTI
SRDNAKNTVYLQMNSLKSEDSAVYYCNARRLFTN
YWGQGTQVTVSP
121 NBX0845 QVQLQESGGGLVQAGGSLRLSCAASGRTFEATYM PirA
GWFRQSPGKEREFVAAISWGGGTTYYGDSVKGRFT
VSRDNAKNTAYLQMNSLKLEDTAVYSCAAATVD
WGPRGPYRYWGQGTQVTVSS
122 NBX0846 QVQLQESGGGLVQAGGSLRLSCAASGRTFSTYYKG PirA
WFRQAPGKEREFLAAISDGGTYYADSVKGRFTISR
DNAKNTVYLQMNSLKPEDTAVYYCAAQGVWRGT
WO 2020/008254 PCT/1B2019/000687
17

GSYTWQYSYDYWGQGTQVTVSS
NBX0849 QVQLQESGGGLVQAGGSLRLSCAASGRTFNRYAM PirA
GWFRQAPGKEREFVAAISWSGNTQYTDSVKGRFTI
SRDDAKNTVYLQMNSLKPEDTAVYYCALRIPASSS
TTYYYADQYDYWGQGTQVTVSS
124 NBX0850 QVQLQESGGGLVQAGGSLRLSCAASGSISASYVMG PirB
WYRQTPGKQRELVATTTSGGTTRYADSVRGRFTIS
RDNARNTVYLQMNSLKPDDTAVYYCNARRLFLNY
WGQGTQVTVSS
125 NBX0851 QVQLQQSGGGLVQPGGSLRLSCAASGRMFSGYVM PirB
GWYRQAPGKQRELVATITNGGLTNYADSVKGRFTI
SRDNAKNTVYLQMNSLKPDDTAVYYCNARRLWDI
YWGQGTQVTVSS
126 NBX0852 QVQLQESGGGLVQAGGSLRLSCEASGRTFSSYRVG PirB
WFRQAPGKGREFVAAISATGGTTYYGDSVKGRFTI
SRDNAENTVSLQMNSLEPEDTAVYYCAATKGIVNY
RVAGTYDAWGQGTQVTVSS
127 NBX0853 QVQLQESGGGLVQAGGSLRLSCSASGSISSSHVMG PirB
WYRQAPGKQRELVATITNGGLTHYADSVKGRFTIS
RDNAKNTVYLQMNSLKPDDTAVYYCNARRLWDN
YWGQGTQVTVSS
128 NBX0854 QVQLQQSGGGLVQAGGSLRISCAASGSISSAYVMG PirB
WYRQAPGTQRELVATITSGGTTNYADSVKGRFTVS
RDNAKNTVYLQMNSLKPDDTAVYYCNARRLWTT
YWGQGTQVTVSS
129 NBX0855 QVQLQESGGGLVQPGESLRLSCAASTSGFSSYVMA PirB
WYRQAPGKQRELVASMTTGGLTNYADSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAAYYCNARRLWN
AYWGQGTQVTVSS
130 NBX0856 QVQLQESGGGLVQAGGSLRLSCVSSGSISASHVMG PirB
WYRQAPGKQRELVATITSGGTTRYADSVKGRFTVS
RDNAKNTVYLQMNDLKSEDTAVYYCHARRLWDT
YWGQGTQVTVSS
131 NBX0857 QVQLQQSGGGLVQPGGSLRLSCAASGRIFSGHVMG PirB
WYRQAPGKQRELVATITNGGLTNYGDSVKGRFTIS
RDNAKNTVYLQMNSLKPDDTAVYYCNARRLWDT
YWGQGTQVTVSS
132 NBX0858 QVQLQESGGGLVQAGGSLRLSCAASRRDFTTTTMA PirB
WYRQAPGKKRETVATVNTGGLTWYADFVKGRFTI
SRDDAVNTLLLQMDSLKPEDTAVYYCNAVLVRAR
GMWGQGTQVTVSS
GWYRQGPGKQRELVATVTSGGMTHYGDSVKGRFT
ISRDNAKNTVYLHMNSLKPEDTGVYYCFARRLWDI
HWGQGTQVTVSS
134 NBX0860 QVQLQESGGGLAQAGGSLGLSCAASETIDSSHVMG PirB
WYRQAPGKQRELVATITSGGLTNYADSAKGRFTIS
RDNAKNAVYLQMNSLKPEDTAVYYCHARRLFRVY
WO 2020/008254 PCT/1B2019/000687
18

SEQ
WGQGTQVTVSS
135 NBX0861 QVQLQESGGGLVQAGGSLRLSCVASGSISNSHVMG PirB
WYRQAPGKERELVATLTSGGLTHFGDSVKGRFTIS
RDNAKNTIYLQMNSLKVEDTAVYYCNARRLLTSM
WGQGTQVTVSP
136 NBX0862 QVQLQESGGGLVQPGGSLRLSCSASGNRFSSSYVL PirB
GWYRQAPGKQRELVATVTSGGLTHYGDSVKGRFT
ISRDNAKNTVYLQMNSLKPEDTAVYYCNARILLTN
YWGQGTQVTVSS
137 NBX0863 QVQLQQSGGGLAQAGGSLRLSCAASGRTFNWYTM PirB
AWFRQAPGKEREFVAAIRLGSTVYGDSVKARFTISR
DNAKSTVSLQMNSLKPEDTALYYCAVGITGDGTIQ
GGPYQYWGQGTQVTVSS
138 NBX0864 QVQLQESGGGLVQPGGSLRLSCAASGIISSAYVMG PirB
WYRQAPGKQRELVATITSGGTTRYADSVKGRITISR
DNAKNTVLLQMNSLKPEDTAVYYCNARILLTNYW
GQGTQVTVSS
139 NBX0865 QVQLQESGGGLVQAGGSLRLSCADSGRVFSTYVM PirB
GWYRQVPGKQRELVATITPGGLINYGDAVKGRFTI
SRDNAKNTVYLQMNSLKPADTAVYYCNARRLFAI
NWGGGTQVTVSS
140 NBX09001 QVQLQESGGGSVQPGGSLRLSCAASGSALSSNVLG PirB
WYRQAPGKQRELVATISSGGLTNYADSVKGRFTIS
RDNAKNTVYLQMNSLKPEDTAVYYCQSRRLFTVY
WGQGTQVTVSS
141 NBX09002 QVQLQESGGGLVQAGESLRLSCADSGRTSSTYDMA PirB
WFRQAPGKEREFVAAISRDGGRLSYADSVKGRFTIS
RDNAKNTLSLQMNSLRPEDTAVY YCAAFSIRGGLR
PSYKYWGQGTQVTVSS
142 NBX09003 QVQLQESGGGLVQPGGSLRLSCTASGSIFSLLNMG PirB
WYRQAPGKQRELVASITSRSYTNYADSVKGRFTISR
DNTKNMVYLQMNSLKPEDTAVYYCNLNPADWGR
LRNWGQGTQVTVSS
143 NBX09004 QVQLQESGGGVVQSGGSLRLSCAGPRSIFSGNAMA PirB
WYRQAPGKQRETVATVNTGGLTWY VDFVKGRFTI
SRDDAKNTLLLQMDSLKPEDTAVYYCNAVLVRAR
GMWGQGTQVTVSS
144 NBX09005 QVQLQESGGGLVQAGGSLRLSCAASGLTFGSYAM PirB
GWFRQAPGKEREFVAAIMRYSSRTYYTDSVKGRFT
ISRDNAKNTVNLQMNNLEPEDTAIYYCAAAKRLSI
VTLPRQYEFWGQGTQVTVSS
145 NBX09006 QVQLQESGGGLVQPGGSLRLSCAASGSISGSYVMG PirB
WYRQAPGKQRELVATITSGGLTRYADSVKGRWTIS
RDNAKNTVYLQMNNLKLEDTAVYYCSARRIATTY
WGQGTQVTVSS
146 NBX09007 QVQLQESGGGLVQPGGSLRLSCAASGSISSSYVMG PirB
WYRQAPGKQRDLVATITNAGNIHYGDSVKGRFTIS
RDNAKNTVSLQMNSLKPEDTAVYYCNARALWRA
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SEQ
YWGQGTQVTVSS
147 NBX09008 | QVQLQESGGGLVQAGGSLRLSCSASGRTFSVRAMG PirB
WFRQAPGKERESVAATHQNTRTTLYADSVKGRFAI
SRDGTKNTVYLQMNSLKPEDTAVYYCAASDDYGL
QIKEVAYKYWGQGTQVTVAS
148 NBX09009 | QVQLQESGGGLVQAGGSLRLSCAASGLTFGSYAM PirB
GWFRQAPGKEREFVATIMRYSSRTYYTDSVKGRFT
ISRDNAKNTVNLQMNNLEPEDAAIYYCAAAKRLSI
VALPRQYEFWGQGTQVTVSS
149 NBX09010 | QVQLQQSGGGLVQAGGSLRLSCAASGLTFGSYAM PirB
GWFRQAPGKEREFVAAIMRYSSRTYYTDSVKGRFT
ISRDNAKNTVNLQMNNLEPEDTAIYYCAAAKRLSR
VTLPREYEFWGQGTQVTVSS
150 NBX09011 | QVQLQESGGGLVQPGESLRLSCAASTSGFSSYVMA PirB
WYRQAPGKQRELVASMTTGGLTNYADSVKGRFTI
SRDNAKNTVYLQMNSLKPEDTAAYYCNARRLWN
AYWGQGTQVTVSS
Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

CDR1 CDR2 CDR3 CDR3
Amino CDR1 Amino CDR2 Amino SEQ
Acid SEQ Acid SEQ ID Acid D
NBX Sequence | ID NO: | Sequence NO: Sequence NO: |Antigen
NBX0401 | GRTFSS |7 IDWWSS |13 AASGKYG | 19 PirA
YT™M Ss LAYSRRD
YAY
NBX0402 | GLPFINY |8 IDWNGD | 14 ASHYQPYI | 20 PirA
AM ST RVSATRR
FEADY
NBX0403 | GLPFINY |9 IDWNGD | 15 AADYQPY |21 PirA
AM ST IRVSATRR
FEADY
NBX0404 | GRTFSR | 10 ISWSGT | 16 ASGSRRL |22 PirB
YTM YYSSDIDY
NBX0405 | GRTFSR | 11 ISWSGT |17 VVGSRRL |23 PirB
YTM YYSSDINY
NBX0406 | GFTFSTY | 12 ISRDGIT | 18 AVVKEDN | 24 VP24
™ RYWCHA
DRNLYRN
NBX0601 | RFTFSTY | 151 ISVGGGI | 273 AKGGKTS | 395 PirA
PM K YTRE
NBX0602 | GRTFSR | 152 VDWSG | 274 AARARDV | 396 PirA
YAM GST YGRAWY
VEDSSTY
DY
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CDR1 CDR2 CDR3 CDR3
Acid SEQ Acid SEQ ID Acid ID
NBX0603 | GSMFSIN | 153 ISRGGIT | 275 NAENRRL | 397 PirA
AM GDDF
NBX0604 | GRTFSS | 154 IRWSGG | 276 AADRDGY | 398 PirA
YT™M SR SSYAHQY
DY
NBX0605 | GSFFSIY | 155 ITSGSET | 277 NRWAPLT | 399 PirA
AM RLDY
NBX0606 | GRTSSSF | 156 ITRTGGR | 278 AARWAT | 400 PirA
AM T ATSNSIRV
YYNEGQY
DY
NBX0607 | GGTFSR | 157 VSWVAE | 279 AAGPRDM | 401 PirA
LT™M TT IRSRNIRS
YDS
NBX0608 | GSMFNI | 158 ISSGGIT | 280 NAENRRL | 402 PirA
NAM GDDY
NBX0609 | GSIFSGN | 159 ISSGGIT | 281 NLENRRL | 403 PirA
VM GDDY
NBX0610 | GSIFSRD | 160 ISSGGIT | 282 NAENRRL | 404 PirA
AM GDDY
LT™M TT IRSRNIKS
YDS
NBX0612 | GTIFSIN | 162 ARSGGII | 284 NALIHTR | 406 PirA
KM YDRVTGY
NBX0613 | GSIFSIN | 163 ISSGGIT | 285 NAENPRL | 407 PirA
AM GDDYW
NBX0614 | GSSFRSN | 164 ITRSGST | 286 HDETMKL | 408 PirB
Al ISVKNDY
NBX0615 | GSMFSR | 165 ISSGGIT | 287 NAENRRL | 409 PirA
NAM GDDY
NBX0616 | GLTFSSY | 166 ISWSGK | 288 AADYQRL | 410 PirA
AM ST GLLRVGV
AEYDY
NBX0617 | GRSGST | 167 IRWSSG | 289 AADNYPL | 411 PirA
SFM MT HIGHQDH
EVDY
NBX0618 | GSIFSFN | 168 ITKGGST | 290 NVKTLRS | 412 PirA
AM ALFPGYE
Y
LT™M TT IRSRNIRS
YDS
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CDR1 CDR2 CDR3 CDR3
LTL MT VRSRNIRS
YDS
NBX0621 | GRTFST | 171 IRWSGG | 293 AADRDGY | 415 PirA
YSM SR SSYAHQY
DY
NBX0622 | GRLENIN | 172 ISSGGIT | 294 NAENRGL | 416 PirA
AM GDDY
NBX0623 | GRTLSN | 173 ISRSGMS | 295 AARGGLP | 417 PirA
YAM T NPSRTYG
FEEQYDY
NBX0624 | GRTVSS | 174 LNWSGT | 296 AAKGAIY |[418 PirA
LPM ST YSYSPRN
NNNYDV
NBX0625 | GSMFNI | 175 ISSGGIT | 297 NAENRLG | 419 PirA
NAM DDY
NBX0626 | ETTLAT | 176 ISSVSSG | 298 NGVRRGR | 420 PirB
NAM GIT SY
NBX0627 | GSSFRSN | 177 ITRSGST | 299 HDETMKL | 421 PirB
Al ITGKNDY
NBX0628 | GRTFSS | 178 IKWSGG | 300 AADRDGY | 422 PirA
YAM SR SRYAHQY
DY
LTI TT IRSRNIRS
YDS
NBX0630 | GVTFSD | 180 ISAGGG | 302 NLKTSYF | 424 PirA
YPM LI WPW
NBX0631 | GGTFSR | 181 VSWVA | 303 AAGPQDM | 425 PirA
LTI QTT IRSRNIRS
YIS
NBX0632 | GGTFSR | 182 VSWVA | 304 AAGPRDM | 426 PirA
LT™M GTT IRSRNIRS
YDS
NBX0633 | GTIFRSK | 183 ISGLGHT | 305 NTFTAAFS | 427 PirA
SM
NBX0634 | GGSFSR | 184 VSWVAE | 306 AAGPRDM | 428 PirA
LTL TT IRSRNIRS
YAS
NBX0635 | GLTFSN | 185 ISWSGK | 307 AAEYQRL | 429 PirA
YAM ST GLLRDGV
ADYSY
LT™M TT IRSRNIRS
YVS
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CDR1 CDR2 CDR3 CDR3
NBX0637 | GRTFSS | 187 ISAGGG | 309 NLKTSYF | 431 PirA
YPM LI WP
NBX0638 | GSIFSGN | 188 ITGGGIT | 310 NYRRIMQ | 432 PirB
VM QY
NBX0639 | GIVFSSI | 189 ITNGGL | 311 NARRIMT | 433 PirB
VM v SY
NBX0640 | GSIFSGN | 190 ITGGGIT | 312 HYRRIMQ | 434 PirB
VM QY
NBX0641 | GSISNSY | 191 ITSGGLT | 313 NARVIFTT | 435 PirB
VM Y
NBX0642 | TVTFSR | 192 ISGSGHR | 314 AGDLVAK | 436 PirB
YAM T FDSAYRV
SYDS
NBX0643 | GSIFSGN | 193 MTGGG | 315 HYRRIMQ | 437 PirB
VM VT QY
NBX0644 | GSIFSGN | 194 ITGGGIT | 316 FYRRIMQ | 438 PirB
VM QY
NBX0645 | GSIFSGN | 195 ITGGGIT | 317 LYRRIMQ | 439 PirB
VM QY
NBX0646 | GSIFSGN | 196 ITGGGV | 318 HYRRIMQ | 440 PirB
VM T Qs
NBX0647 | GSIFSGN | 197 MTGGG | 319 HYRRIMQ | 441 PirB
VM VT QY
NBX0648 | GIIFSSN | 198 RTSGGL | 320 NARRLFT | 442 PirB
VM T NY
NBX0649 | GSIASGN | 199 ITSGGLT | 321 NYRRLAT | 443 PirB
VL GY
NBX0650 | GSIFSGN | 200 ITGGGIT | 322 HYRRIMQ | 444 PirB
VL QY
NBX0722 | GRTFSS | 201 ISRSGDP | 323 AARQIYS | 445 VP28
YPM G NNYSY
NBX0723 | GRTFSS | 202 ISRSGVP | 324 AARSIYSN | 446 VP28
YPM G NYSY
NBX0724 | GRTFSS | 203 ISRSGVS | 325 AARSIYSN | 447 VP28
YPM G NYSY
NBX0725 | GRTFSS | 204 ISRSGNP | 326 AARQIYS | 448 VP28
YPM G NNYSY
NBX0730 | GRTFSS | 205 INRNGNI | 327 AARTIYDS | 449 VP28
YPM P HYTS
NBX0737 | DGAITG | 206 ITYDGST | 328 ARAGEEY | 450 VP24
SYYVW ICSGYGC
HGSLGLD
Y
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CDR1 CDR2 CDR3 CDR3
NBX0738 | GFTFGS | 207 INSGDG | 329 ATGIRTPII | 451 VP24
YAM RI
NBX0739 | GSIFTSD | 208 MTSAGT | 330 GVGRF 452 VP24
v T
NBX0745 | GRTFSS | 209 ISRSGDP | 331 AAREIYSN | 453 VP28
YPM G NYSY
NBX0746 | GRTFSS | 210 ISRSGNP | 332 AARQIYS | 454 VP28
YAM G NNYSY
NBX0813 | GMLFSIR | 211 IGSSGNT | 333 NALNY 455 PirA
NM
NBX0814 | MRTFNS | 212 TAWTGG | 334 AAQTRPY | 456 PirA
RTI N DLPSIRPD
DYAS
NBX0815 | GRTFSS | 213 VSWSGG | 335 AAQRVM | 457 PirA
YAM ST DYYRPRT
ESAYAY
NBX0816 | EYIFSNF | 214 ISRSGSR | 336 AAVYGQY | 458 PirA
GM M SYHYSSD
SKQYSY
NBX0817 | GGTNSN | 215 LSWSGY | 337 AARLSGR | 459 PirA
YAM NT TAGSRTY
YAEGQYD
Y
NBX0818 | GRTSSSS | 216 IRWSDG | 338 AAATTDW | 460 PirA
YL ST GPRGPYN
Y
NBX0819 | GRTSGA | 217 ISGSGRT | 339 AASTFDW | 461 PirA
WNM T GPRGPYR
L
NBX0820 | GRTFSN | 218 VGRGIN | 340 AVTSRWG | 462 PirA
YVI SAYHAT QFDRTDF
NSW
NBX0821 | GMLFSIR | 219 IGTSGAT | 341 NALNY 463 PirA
NM
NBX0822 | GQTFTN | 220 VSRSGL | 342 AATTLDW | 464 PirA
SGM GR GPRGPYR
Y
NBXO0823 | GSIFSIDF | 221 IRGGNT | 343 NAQITMR | 465 PirA
M GGTWSTS
EY
NBX0824 | GRTLSS | 222 ITWNGD | 344 AADTVGR | 466 PirA
YLSSYA RT WRSTLSV
M RDEYDY
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CDR1 CDR2 CDR3 CDR3
NBX0825 | GRTFSA | 223 MSRGSA | 345 AGGSPGS | 467 PirA
YT™M A SQIATPEA
YTY
NBX0826 | GSISSSH | 224 ITSGGST | 346 HARRLWN | 468 PirB
VM TY
NBXO0827 | GSISSSF | 225 ITSGGST | 347 HARRLWN | 469 PirB
VM TY
NBX0828 | RSIFSGN | 226 VNTGGL | 348 NAVLVRA | 470 PirB
AM T RGM
NBX0829 | GDRLSS | 227 VTSGGR | 349 NARILFTN | 471 PirB
YVM T Y
NBX0830 | GSVLSR | 228 ITSGGIT | 350 NARALW | 472 PirB
YVM NTY
NBX0831 | GDAFSR | 229 ITSGGST | 351 HARRLWN | 473 PirB
YVM TY
NBX0832 | GSISSSY | 230 ITSGGST | 352 HARRLWN | 474 PirB
VM TY
NBXO0833 | GSISSSH | 231 ITSGGST | 353 HARRLWD | 475 PirB
VM TY
NBX0834 | ESMFRD | 232 ISRGGLI | 354 NARRLLT | 476 PirB
HNM vV
NBXO0835 | GNRFSSS | 233 VTSGGL | 355 NARILLTN | 477 PirB
YVM T Y
NBX0836 | GIRLGSY | 234 VTSGGT | 356 NARILFTN | 478 PirB
VM T Y
NBXO0837 | GIVFAN | 235 ITNGGLI | 357 NARRLYQ | 479 PirB
HVM QY
NBX0838 | GRTVGS | 236 IGWSGA | 358 AQRPSSR | 480 PirB
YAM ST YASRYLG
DYAY
NBX0839 | GSIGSDY | 237 ITSGGLT | 359 NARRLFR | 481 PirB
VL NY
NBX0840 | GSIRSSN | 238 MTAGGL | 360 HYRRIFN | 482 PirB
VM T VY
NBX0841 | ARTFIYK | 239 IMWSVG | 361 AAATTST | 483 PirB
M NNY QWRY
NBX0842 | GSIDNG | 240 ITSGTNT | 362 LARRLFT | 484 PirB
YVM MY
NBX0843 | GNRFSSS | 241 VTTGGL | 363 NARILLTN | 485 PirB
YVM T Y
NBX0844 | GRMFSG | 242 ITNGGLT | 364 NARRLFT | 486 PirB
FVM NY
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CDR1 CDR2 CDR3 CDR3
NBX0845 | GRTFEA | 243 ISWGGG | 365 AAATVD | 487 PirA
TYM TT WGPRGPY
RY
NBX0846 | GRTEST | 244 ISDGGT | 366 AAQGVW | 488 PirA
YYK RGTGSYT
WQYSYD
Y
NBX0849 | GRTENR | 245 ISWSGN | 367 ALRIPASS | 489 PirA
YAM T STTYYYA
DQYDY
NBX0850 | GSISASY | 246 TTSGGT | 368 NARRLFL | 490 PirB
VM T NY
NBX0851 | GRMFSG | 247 ITNGGLT | 369 NARRLWD | 491 PirB
YVM Iy
NBX0852 | GRTFSS | 248 ISATGGT | 370 AATKGIV | 492 PirB
YRV T NYRVAGT
YDA
NBXO0853 | GSISSSH | 249 ITNGGLT | 371 NARRLWD | 493 PirB
VM H NY
NBX0854 | GSISSAY | 250 ITSGGTT | 372 NARRLWT | 494 PirB
VM TY
NBXO0855 | TSGFSSY | 251 MTTGGL | 373 NARRLWN | 495 PirB
VM T AY
NBX0856 | GSISASH | 252 ITSGGTT | 374 HARRLWD | 496 PirB
VM TY
NBXO0857 | GRIFSGH | 253 ITNGGLT | 375 NARRLWD | 497 PirB
VM TYW
NBX0858 | RRDFTT | 254 VNTGGL | 376 NAVLVRA | 498 PirB
TTM T RGM
NBXO0859 | GNRFSSS | 255 VTSGGM | 377 FARRLWD | 499 PirB
YVM T H
NBX0860 | ETIDSSH | 256 ITSGGLT | 378 HARRLFR | 500 PirB
VM VY
NBX0861 | GSISNSH | 257 LTSGGL |379 NARRLLT | 501 PirB
VM T SM
NBX0862 | GNRFSSS | 258 VTSGGL | 380 NARILLTN | 502 PirB
YVL T Y
NBX0863 | GRTFNW | 259 IRLGST | 381 AVGITGD | 503 PirB
YT™M GTIQGGP
YQY
NBX0864 | GIISSAY | 260 ITSGGTT | 382 NARILLTN | 504 PirB
VM Y
NBX0865 | GRVFST | 261 ITPGGLI | 383 NARRLFALI | 505 PirB
YVM N
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CDR1 CDR2 CDR3 CDR3
NBX0900 | GSALSS | 262 ISSGGLT | 384 QSRRLFT | 506 PirB
1 NVL VY
NBX0900 | GRTSST | 263 ISRDGG | 385 AAFSIRGG | 507 PirB
2 YDM RL LRPSYKY
NBX0900 | GSIFSLL | 264 ITSRSYT | 386 NLNPADW | 508 PirB
3 NM GRLRN
NBX0900 | RSIFSGN | 265 VNTGGL | 387 NAVLVRA | 509 PirB
4 AM T RGM
NBX0900 | GLTFGS | 266 IMRYSS | 388 AAAKRLSI | 510 PirB
5 YAM RT VTLPRQY
EF
NBX0900 | GSISGSY | 267 ITSGGLT | 389 SARRIATT | 511 PirB
6 VM Y
NBX0900 | GSISSSY | 268 ITNAGNI | 390 NARALWR | 512 PirB
7 VM AY
NBX0900 | GRTESV | 269 IHQNTR | 391 AASDDYG | 513 PirB
8 RAM TT LQIKEVA
YKY
NBX0900 | GLTFGS | 270 IMRYSS | 392 AAAKRLSI | 514 PirB
9 YAM RT VALPRQY
EF
NBX0901 | GLTFGS | 271 IMRYSS | 393 AAAKRLS | 515 PirB
0 YAM RT RVTLPRE
YEF
NBX0901 | TSGFSSY | 272 MTTGGL | 394 NARRLWN | 516 PirB
1 VM T AY
Antibodies recombinantly expressed

[0037] In another aspect, the present invention provides a method for producing VyH in a
suitable producing organism. Suitable producing organisms include, without limitation, bacteria,
yeast and algae. In certain embodiments, the producing bacterium is Escherichia coli. In certain
embodiments, the producing bacterium is a member of the Bacillus genus. In certain
embodiments, the producing bacterium is a probiotic. In certain embodiments, the yeast is
Pichia pastoris. In certain embodiments, the yeast is Saccharomyces cerevisiae. In certain
embodiments, the algae is a member of the Chlamydomonas or Phaeodactylum genera.
Antibodies added to feed
[0038] In yet another aspect, the present invention provides a polypeptide or pluralities thereof
comprising a VygH or VyHs that bind disease-causing agents and are administered to host
animals via any suitable route as part of a feed product. In certain embodiments, the animal is
WO 2020/008254 PCT/IB2019/000687
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selected from the list of host animals described, with that list being representative but not
limiting. In certain embodiments, the route of administration to a recipient animal can be, but is
not limited to: introduction to the alimentary canal orally or rectally, provided to the exterior
surface (for example, as a spray or submersion), provided to the medium in which the animal
dwells (including air and water based media), provided by injection, provided intravenously,
provided via the respiratory system, provided via diffusion, provided via absorption by the
endothelium or epithelium, or provided via a secondary organism such as a yeast, bacterium,
algae, bacteriophages, plants and insects. In certain embodiments, the host animal is a shellfish.
In certain embodiments, the host animal is shrimp
Feed product
[0039] In a further aspect, the present invention provides a polypeptide or pluralities thereof
comprising a VyzH or VyHs that bind disease-causing agents and are administered to host
animals in the form of a product. The form of the product is not limited, so long as it retains
binding to the disease-causing agent in the desired form. In certain embodiments, the product is
feed, pellet, nutritional supplement, premix, therapeutic, medicine, or feed additive, but is not
limited to these forms.

Feeding dosage
comprising a VyzH or VyHs that bind disease-causing agents and are administered to host
animals as part of a product at any suitable dosage regime. In practice, the suitable dosage is the
dosage at which the product offers any degree of protection against a disease-causing agent, and
depends on the delivery method, delivery schedule, the environment of the recipient animal, the
size of the recipient animal, the age of the recipient animal and the health condition of the
recipient animal among other factors. In certain embodiments, VyHs are administered to
recipient animals at a concentration in excess of 1 mg/kg of body weight. In certain
embodiments, VyHs are administered to recipient animals at a concentration in excess of 5
mg/kg of body weight. In certain embodiments, VyHs are administered to recipient animals at a
concentration in excess of 10 mg/kg of body weight. In certain embodiments, VyHs are
administered to recipient animals at a concentration in excess of 50 mg/kg of body weight. In
certain embodiments, ViHs are administered to recipient animals at a concentration in excess of
100 mg/kg of body weight. In certain embodiments, VyHs are administered to recipient animals
at a concentration less than 1 mg/kg of body weight. In certain embodiments, VyHs are
administered to recipient animals at a concentration less than 500 mg/kg of body weight. In
certain embodiments, ViyHs are administered to recipient animals at a concentration less than
100 mg/kg of body weight. In certain embodiments, VyHs are administered to recipient animal
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at a concentration less than 50 mg/kg of body weight. In certain embodiments, VyHs are
administered to recipient animals at a concentration less than 10 mg/kg of body weight.
Feeding frequency
comprising a VygH or VyHs that bind disease-causing agents and are administered to host
animals as part of a product at any suitable dosage frequency. In practice, the suitable dosage
frequency is that at which the product offers any protection against a disease-causing agent, and
depends on the delivery method, delivery schedule, the environment of the recipient animal, the
size of the recipient animal, the age of the recipient animal and the health condition of the
recipient animal, among other factors. In certain embodiments, the dosage frequency can be but
is not limited to: constantly, at consistent specified frequencies under an hour, hourly, at
specified frequencies throughout a 24-hour cycle, daily, at specified frequencies throughout a
week, weekly, at specified frequencies throughout a month, monthly, at specified frequencies
throughout a year, annually, and at any other specified frequency greater than 1 year.
Feed additives
comprising a VgH or ViHs that bind disease-causing agents and are administered to host
animals as part of a product that also comprises other additives or coatings. In practice, the most
suitable coating or additive depends on the method of delivery, the recipient animal, the
environment of the recipient, the dietary requirements of the recipient animal, the frequency of
delivery, the age of the recipient animal, the size of the recipient animal, the health condition of
the recipient animal In certain embodiments, these additives and coatings can include, but are
not limited to the following list and mixtures thereof: a vitamin, an antibiotic, a hormone, 1
peptide, a steroid, a probiotic, a bacteriophage, chitin, chitosan, B-1,3-glucan, vegetable extracts,
peptone, shrimp meal, krill, algae, B-cyclodextran, alginate, gum, tragacanth, pectin and gelatin.
Non-feed uses
comprising a VygH or VyHs that bind disease-causing agents, and can be used in a non-feed use,
such as but not limited to: a diagnostic kit, an ELISA-based assay, a western blot assay, an
immunofluorescence assay, or a FRET assay, in its current form and/or as a polypeptide
conjugated to another molecule. In certain embodiments, the conjugated molecule is can be but
is not limited to: a fluorophore, a chemiluminescent substrate, an antimicrobial peptide, a
nucleic acid or a lipid.
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Antigens
comprising a VyH or VyHs that bind disease-causing agents, including toxins, produced by a
species of Vibrio. In certain embodiments, the Vibrio species is capable of harbouring the pVA-
1 plasmid. In certain embodiments, the species does not belong to the Vibrio genus but is
capable of harbouring disease-causing agents shared by Vibrio species, such as but not limited to
the pVA-1 plasmid. In certain embodiments, the Vibrio species refers to both current and
reclassified organisms. In certain embodiments, the Vibrio species is V. adaptatus, V. aerogenes,
V. aestivus, V. aestuarianus, V. agarivorans, V. albensis, V. alfacsensis, V. alginolyticus, V.
anguillarum, V. areninigrae, V. artabrorum, V. atlanticus, V. atypicus, V. azureus, V.
brasiliensis, V. bubulus, V. calviensis, V. campbellii, V. casei, V. chagasii, V. cholerae, V.
cincinnatiensis, V. coralliilyticus, V. crassostreae, V. cyclitrophicus, V. diabolicus, V.
diazotrophicus, V. ezurae, V. fluvialis, V. fortis, V. furnissii, V. gallicus, V. gazogenes, V.
gigantis, V. halioticoli, V. harveyi, V. hepatarius, V. hippocampi, V. hispanicus, V. ichthyoenteri,

V.indicus, V. kanaloae, V. lentus, V. litoralis, V. logei, V. mediterranei, V. metschnikovii, V.
mimicus, V. mytili, V. natriegens, V. navarrensis, V. neonatus, V. neptunius, V. nereis, V.
nigripulchritudo, V. ordalii, V. orientalis, V. pacinii, V. parahaemolyticus, V. pectenicida, V.
penaeicida, V. pomeroyi, V. ponticus, V. proteolyticus, V. rotiferianus, V. ruber, V. rumoiensis,
V. salmonicida, V. scophthalmi, V. splendidus, V. superstes, V. tapetis, V. tasmaniensis, V.
tubiashii, V. vulnificus, V. wodanis, V. xuii, V. fischer, or V. hollisae
[0045] In certain embodiments, the VyH or plurality thereof is capable of binding to two or
more disease-causing agents, originating from the same or different species. In certain
embodiments, the disease-causing agent is a polypeptide with 60%, 70%, 80%, 90%, 95%, 98%,
99%, or 100% amino acid sequence identity to PirA (SEQ ID 25). In certain embodiments, the
disease-causing agent is a polypeptide with 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%
amino acid sequence identity to PirB (SEQ ID 26). In certain embodiments, the disease-causing
agent is an exposed peptide, protein, protein complex, nucleic acid, lipid, or combination
thereof, that is associated to the surface of the Fibrio bacterium. In certain embodiments, the
disease-causing agent is a pilus, fimbria, flagellum, secretion system or porin. In certain
embodiments, the disease-causing agent is the Vibrio bacterium
comprising a VygH or VyHs that bind disease-causing agents produced by White Spot Syndrome
Virus. In certain embodiments, the disease-causing agent is a polypeptide with 60%, 70% 80%,
90%, 95%, 98%, 99%, or 100% amino acid sequence identity VP24 (SEQ ID 27). In certain
embodiments, the disease-causing agent is a polypeptide with 60%, 70% 80%, 90%, 95%, 98%,
WO 2020/008254 W PCT/IB2019/000687
99%, or 100% amino acid sequence identity to VP28 (SEQ ID 28). In certain embodiments, the
disease-causing agent is viral protein associated with or hypothesised to be associated with the
envelope of the White Spot Syndrome Virus. In certain embodiments, the disease-causing agent
is the White Spot Syndrome Virus.
SEQ ID 25:
PirA
>tr|AOA085YLCO|AOAO085YLCO_VIBPH JHE-like toxin PirA-like OS=Vibrio
parahaemolyticus OX=670 GN=vp19 PE=4 SV=1

MSNNIKHETDY SHDWTVEPNGGVTEVDSKHTPIIPEVGRSVDIENTGRGELTIQYQWGA
PFMAGGWKVAKSHVVQRDETYHLQRPDNAFYHQRIVVINNGASRGFCTIYYH
SEQ ID 26:
PirB
>tr|AOA085YLC1|A0A085YLC1_VIBPH JHE-like toxin PirB-like OS=Vibrio
parahaemolyticus OX=670 GN=BTO19_25780 PE=4 SV=1
MTNEYVVTMSSLTEFNPNNARKSYLFDNYEVDPNYAFKAMVSFGLSNIPYAGGFLSTL
WNIFWPNTPNEPDIENIWEQLRDRIQDLVDESIIDAINGILDSKIKETRDKIQDINETIENFG
YAAAKDDYIGLVTHYLIGLEENFKRELDGDEWLGYAILPLLATTVSLQITYMACGLDY
KDEFGFTDSDVHKLTRNIDKLYDDVSSYITELAAWADNDSYNNANQDNVYDEVMGAR
SWCTVHGFEHMLIW QKIKELKKVDVFVHSNLISY SPAVGFPSGNFNYIATGTEDEIPQPL
KPNMFGERRNRIVKIESWNSIEIHY YNRVGRLKLTYENGEVVELGKAHKYDEHYQSIEL
NGAYIKYVDVIANGPEAIDRIVFHFSDDRTFVVGENSGKPSVRLQLEGHFICGMLADQE
GSDKVAAFSVAYELFHPDEFGTEK
SEQ ID 27:
VP24
>tr/QOE7K6|Q9E7K6_WSSV Major structural protein VP24 OS=White spot syndrome virus

0X=92652 GN=VP24 PE=4 SV=1
MHMWGVYAAILAGLTLILVVISIVVTNIELNKKLDKKDKDAYPVESEIINLTINGVARGN
HFNFVNGTLQTRNYGKVYVAGQGTSDSELVKKKGDIILTSLLGDGDHTLNVNKAESKE
LELYARVYNNTKRDITVDSVSLSPGLNATGREFSANKFVLYFKPTVLKKNRINTLVFGA
TFDEDIDDTNRHYLLSMRFSPGNDLFKVGEK
SEQ ID 28
VP28
>tr|A6ZI33|A6Z133_WSSV Coat protein OS=White spot syndrome virus 0X=92652 GN=VP28

PE=4 SV=1
WO 2020/008254 PCT/1B2019/000687
31
MDLSFTLSVVSAILAITAVIAVFIVIFRYHNTVTKTIETHTGNIETNMDENLRIPVTAEVGS
GYFKMTDVSFDSDTLGKIKIRNGKSDAQMKEEDADLVITPVEGRALEVTVGQNLTFEG
TFKMWNNTSRKINITGMQMVPKINPSKAFVGSSNTSSFTPVSIDEDEVGTFVCGTTFGAP
IAATAGGNLFDMYVHVTYSGTETE
EXAMPLES
[0047] The following illustrative examples are representative of the embodiments of the
applications, systems and methods described herein and are not meant to be limiting in any way.
[0048] While preferred embodiments of the present invention are shown and described herein, it
will be obvious to those skilled in the art that such embodiments are provided by way of
example only. Numerous variations, changes, and substitutions will now occur to those skilled
in the art without departing from the invention. It should be understood that various alternatives
to the embodiments of the invention described herein may be employed in practicing the
invention,
1. Production of antigens
[0049] Recombinant antigens can be purified from an £. coli expression system. For example,
the gene for an antigen can be expressed at 18°C in £. coli BL21 (DE3) cells grown overnight in
autoinducing media (Formedium). Cells are then lysed by sonication in buffer A (250 mM
NaCl, 50 mM CaCl,, 20 mM Imidazole and 10 mM HEPES, pH 7.4) with 12.5 ug/ml DNase I,
and 1X Protease inhibitor cocktail (Bioshop). The lysate is cleared by centrifugation at 22000 x
g for 30 minutes at 4°C , and is then applied to a 5 ml HisTrap HP column (GE Healthcare) pre-
equilibrated with buffer A, washed with ten column volumes of buffer A and eluted with a
gradient of 0% to 60% (vol/vol) buffer B (250 mM NaCl, 50 mM CaCl,, 500 mM imidazole
and10 mM HEPES, pH 7.4). The protein is then dialyzed overnight in the presence of TEV
against buffer C (250 mM NaCl, 10 mM HEPES, pH 7.4 and 5 mM B-mercaptoethanol) at 4°C
The dialyzed protein is applied to a HisTrap HP column (GE Biosciences) pre-equilibrated with
buffer C. 6xHis-tagged TEV and 6xHis-tag are bound to the column and the antigen is collected
in the flowthrough. The sample is dialyzed overnight against buffer D (5 mM NaCl and 10 mM
Tris pH 8.8) and then applied to a 5 ml HiTrap Q HP column (GE Healthcare). The protein is
eluted with a gradient of 0% to 50% (vol/vol) buffer E (1.0 M NaCl and 10 mM Tris pH 8.8).
Lastly, the elution is loaded onto a Superdex 75 Increase 10/300 GL gel filtration column (GE
Healthcare) using buffer F (400 mM NaCl and 20 mM HEPES pH 7.4). The protein sample is
then concentrated to 1 mg/mL using Amicon concentrators with appropriate molecular weight
cut-off (MWCO; Millipore). The purified protein is stored at =80°C.
WO 2020/008254 PCT/1B2019/000687
32
2. Production of NBXs and panning

Llama immunization
[0050] A single llama is immunized with purified disease-causing agents, such as the antigens
listed, which may be accompanied by adjuvants. The llama immunization is performed using
100 pg of each antigen that are pooled and injected for a total of four injections. At the time of
injection, the antigens are thawed, and the volume increased to 1 ml with PBS. The 1 ml
antigen-PBS mixture is then mixed with 1 ml of Complete Freund’s adjuvant (CFA) or
Incomplete Freund’s adjuvant (IFA) for a total of 2 ml. A total of 2 ml is immunized per
injection. Whole llama blood and sera are then collected from the immunized animal on days 0,
28, 49, 70. Sera from days 28, 49 and 70 are then fractionated to separate VyH from
conventional antibodies. ELISA can be used to measure reactivity against target antigens in
polyclonal and VyH-enriched fractions. Lymphocytes are collected from sera taken at days 28,
49, and 70.
Panning
[0051] RNA isolated from purified llama lymphocytes is used to generate cDNA for cloning
into phagemids. The resulting phagemids are used to transform £. coli TG-1 cells to generate a
library of expressed VyH genes. The phagemid library size can be ~2.5 x 10” total transformants
and the estimated number of phagemid containing VyH inserts can be estimated to be ~100%.
High affinity antibodies are then selected by panning against the Vibrio or WSSV antigens used
for llama immunization. At least two rounds of panning are performed and antigen-binding
clones arising from rounds 2 or later are identified using phage ELISA. Antigen-binding clones
are sequenced, grouped according to their CDR regions, and prioritized for soluble expression in
E. coli and antibody purification.
[0052] Figure 2 shows the Phage ELISA results for all antibodies of this disclosure. Black bars
show binding to wells coated with the antigen specified in Tables 1 and 2 dissolved in
phosphate-buffered saline (PBS). Grey bars are negative controls that show binding to wells
coated with PBS only. In all cases binding to the antigen target is at least 50% above binding to
the PBS-coated wells. Panel A shows the results for NBX0401 to NBX0406. Panel B shows the
results for NBX0601 to NBX0630. Panel C shows the results for NBX063 1 to NBX0637,
NBX0813 to NBX0825, NBX0845, NBX0846, and NBX0849. Panel D shows the results for
NBX0638 to NBX0650, and NBX0826 to NBX0844. Panel E shows the results for NBX0850 to
NBX0865, and NBX09001 to NBX09011. Panel F shows the results for NBX0722 to
NBX0725, NBX0730, NBX0738, NBX0739, NBX0745, and NBX0746.

WO 2020/008254 PCT/1B2019/000687
33
Purification of VyHs from E. coli

[0053] TEV protease-cleavable, 6xHis-thioredoxin-NBX fusion proteins are expressed in the
cytoplasm of E. coli grown in autoinducing media (Formedium) for 24 hours at 30°C. Bacteria
are collected by centrifugation, resuspended in buffer A (10 mM HEPES, pH 7.5, 250 mM
NaCl, 20 mM Imidazole) and lysed using sonication. Insoluble material is removed by
centrifugation and the remaining soluble fraction is applied to a HisTrap column (GE
Biosciences) pre-equilibrated with buffer A. The protein is eluted from the column using an
FPLC with a linear gradient between buffer A and buffer B (10 mM HEPES, pH 7.5, 500 mM
NaCl, 500 mM Imidazole). The eluted protein is dialyzed overnight in the presence of TEV
protease to buffer C (10 mM HEPES, pH 7.5, 500 mM NaCl). The dialyzed protein is applied to
a HisTrap column (GE Biosciences) pre-equilibrated with buffer C. 6xHis-tagged TEV and
6xHis-tagged thioredoxin are bound to the column and highly purified NBX is collected in the
flowthrough. NBX proteins are dialyzed overnight to PBS and concentrated to ~10 mg/ml
Purification of VyHs from P. pastoris
[0054] Pichia pastoris strain GS115 with constructs for the expression and secretion of 6xHis-
tagged VyH are grown for 5 days at 300C with daily induction of 0.5% (vol/vol) methanol
Yeast cells are removed by centrifugation and the NBX-containing supernatant is spiked with 10
mM imidazole. The supernatant is applied to a HisTrap column (GE Biosciences) pre-
equilibrated with buffer A (10 mM HEPES, pH 7.5, 500 mM NaCl). The protein is eluted from
the column using an FPLC with a linear gradient between buffer A and buffer B (10 mM
HEPES, pH 7.5, 500 mM NaCl, 500 mM Imidazole). NBX proteins are dialyzed overnight to
PBS and concentrated to ~1.5 mg/ml.
3. Protein Pull-downs
[0055] Approximately 0.1 mg of antigen is incubated with NBX at a 1:5 molar ratio in 200 pl of
binding buffer (10 mM phosphate buffer pH7.4 and 500 mM NaCl) for 30 minutes at room
temperature, and then applied onto a column containing Ni-NTA (nickel-nitrilotriacetic acid}
resin pre-equilibrated with the binding buffer. Protein mixture and the resin are incubated for 30
minutes before the resin is washed with the binding buffer and then with the binding buffer plus
20 mM Imidazole. Bound proteins are eluted with 100 pl of 1 M imidazole, pH 7.4. The
presence or absence of NBX in the various fractions is analyzed on an SDS-PAGE gel. A
protein solution containing only the NBX is also applied to a separate column to assess non-
specific binding of the NBX to the resin
[0056] Figure 3 shows representative results for four unique NBXs. For each of the four
antibodies shown, the lanes are as follows. (1) Starting material of PirA(*) and NBX(") mixture
prior to application to Ni-NTA resin. (2) Flow-through of PirA and NBX through the Ni-NTA
WO 2020/008254 o PCT/IB2019/000687
resin. (3) Final wash of the Ni-NTA resin prior to protein elution. (4) Elution of PirA and NBX
from the Ni-NTA resin. (5) Elution from Ni-NTA resin to which only NBX was applied. (6)
Final wash of Ni-NTA resin to which only NBX was applied. (7) NBX(") only mixture prior to
application to Ni-NTA resin. NBXs that can successfully be pulled down by PirA are those that
appear in the lane 4 elution but not in the lane 5 elution. For each gel a ladder of proteins of
known sizes in kilodaltons (kDa) are shown for reference.
4. Protein stability in shrimp intestinal tract fluid
[0057] Thaw frozen shrimp midgut extract and NBX at room temperature, and immediately
place on ice. Spin shrimp midgut extract and protein at 10,000 RCF for 1 minute to pellet and
remove any precipitation. Prechill PBS and saline on ice. Label and prechill 8 x 0.2 mL strip
tubes on ice. Set up two reactions in volumes of 10 pl on ice. The first reaction contains no
shrimp midgut extract and consists of 5 ug NBX in 3.2 L PBS and 4.8 pL of 150 mM NaCl.
The second reaction contains shrimp midgut extract and is generated using the following ratios:
2.4 uL shrimp midgut extract, 5 ug NBX in 0.8 pL PBS, and 4.8 uL of 150 mM NaCl. The
tubes are incubated on ice for 5 minutes (corresponds to time = 0 minutes in Figure 1) followed
by 26°C for up to 24 hours. The final incubation temperature (26°C) is the internal temperature
of a shrimp. After incubation, add 8 pL of preheated 2X SDS sample buffer to stop the reaction.
Boil at 95-100°C for 5 minutes. The stability of each NBXs is assessed by the presence or
absence of the NBX on an 18% SDS-PAGE gel.
[0058] Figure 4 shows representative results for four unique NBXs. For each of the four
antibodies shown SDS-PAGE gels are arranged from left to right as follows. A ladder of
proteins of known sizes in kilodaltons (kDa) are shown for reference. The next two lanes show
the NBX at the beginning and end of the experiment in the absence of shrimp midgut extract.
These lanes show that the NBX is not degraded over time in the absence of shrimp midgut
extract. The subsequent lane shows the appearance of the shrimp midgut extract at the start of
the experiment without NBX added. This lane allows for the visualization of naturally occurring
proteins in the extract. The subsequent 7-9 lanes show the time course of NBX stability in the
shrimp midgut extract. These lanes allow for the visualization of the relative stability of the
NBX. The longer the full-sized NBX can be visualized on the gel the more stable it is. The final
lane shows the shrimp midgut extract in the absence of NBX at the endpoint of the assay.
[0059] All publications, patent applications, issued patents, and other documents referred to in
this specification are herein incorporated by reference as if each individual publication, patent
application, issued patent, or other document is specifically and individually indicated to be
incorporated by reference in its entirety. Definitions that are contained in text incorporated by
reference are excluded to the extent that they contradict definitions in this disclosure.
WO 2020/008254 s PCT/IB2019/000687
[0060] The following references are incorporated by reference in their entirety

1. Kierath, D. (2015, March). The growth of global aquaculture — Fishy business.
Retrieved from deloitte.com/au/en/pages/consumer-business/articles/the-growth-of-aqua-
culture-fishy-business
2. Stentiford, G. D., Sritunyalucksana, K., Flegel, T. W., Williams, B. A. P.,
Withyachumnarnkul, B., Itathitphaisarn, O., Bass, D. (2017). New paradigms to help solve the
global aquaculture disease crisis. PLos Pathogens, 13(2), pp. 1-6
3.Lee, C. T, Chen, T. I, Yang, Y. T, Ko, T. P,, Huang, Y. T., Huang,, J. Y., Huang, M.
F., Lin, S.J, Chen, C. Y, Lin, S. S, Lightner, D. V., Wang, H. C., Wang, A. H. J., Wang, H. C.,
Hor, L. I, Lo, C. F. (2015) The opportunistic marine pathogen Vibrio parahaemolyticus

becomes virulent by acquiring a plasmid that expresses a deadly toxin. PNAS, 112(34), pp
10789-10803.
4. FAO Fisheries and Aquaculture (2013) Report of the FAO/MARD Technical

‘Workshop on Early Mortality Syndrome (EMS) or Acute Hepatopancreatic Necrosis Syndrome
(AHPNS) of Cultured Shrimp (under TCP/VIE/3304). rep. no. 1053. Retrieved from
www.fao.org/docrep/018/i3422¢/i3422e.pdf
5. Tran, L., Numan, L., Redman, R. M., Mohney, L. L., Pantoja, C. R., Fitzsimmons, K.,
Lightner, D. V. (2013) Determination of the infectious nature of the agent of acute

hepatopancreatic necrosis syndrome affecting penaeid shrimp. Diseases of Aquatic Organisms,
105(1), pp. 45-55
6. Ahmed, H. A, El Bayomi, R. M., Hussein, M. A, Khedr, M. H. E., Abo Ramela, E.
M., El-Ashram, A. M. M. (2018) Molecular characterization, antibiotic resistance pattern and

biofilm formation of Vibrio parahaemolyticus and V. cholerae isolated from crustaceans and
humans. International Journal of Food Microbiology, 274, pp. 31-37
7. Flegel, T. (2012) Historic emergence, impact and current status of shrimp pathogens
in Asia, Journal of Invertebrate Pathology, 110(2), pp. 166-173.
8. Lightner, D. V., Redman, R. M., Pantoja, C. R., Tang, K. F. J., Noble, B. L.,
Schofield, P., Mohney, L. L., Nunan, L. M., Navarro, S. A. (2012) Historic emergence, impact
and current status of shrimp pathogens in the Americas, Journal of Invertebrate Pathology, 110
(2), pp. 174-183
[0061] While preferred embodiments of the present invention have been shown and described
herein, it will be obvious to those skilled in the art that such embodiments are provided by way
of example only. Numerous variations, changes, and substitutions will now occur to those
skilled in the art without departing from the invention. It should be understood that various
alternatives to the embodiments of the invention described herein may be employed in practicing
WO 2020/008254 PCT/IB2019/000687
36
the invention. It is intended that the following claims define the scope of the invention and that
methods and structures within the scope of these claims and their equivalents be covered
thereby
WO 2020/008254 PCT/1B2019/000687
37
CLAIMS
WHAT IS CLAIMED IS:
1. A polypeptide comprising at least one variable region fragment of a heavy chain

antibody (VyH), wherein the at least one ViyH specifically binds a species of Vibrio or a White
Spot Syndrome virus.
2 A polypeptide comprising at least one variable region fragment of a heavy chain
antibody (VyH), wherein at least one ViH specifically binds an aquaculture associated
pathogen.
3. The polypeptide of claim 1 or 2, wherein the VgH comprises an amino acid
sequence with at least 80% identity to the amino acid sequence set forth in any one of SEQ ID
Nos: 1 or2or4or5or53or9 or97or 121
4. The polypeptide of claim 1 or 2 or 3, wherein the polypeptide comprises a
plurality of VyHs.
5 The polypeptide of claim 4, wherein the polypeptide comprises at least three
VuHs.
6. The polypeptide of claim 4 or 5, wherein any one of the plurality of VyHs is

identical to another VyH of the plurality of ViHs.
7. The polypeptide of any one of claims 4 to 6, wherein the plurality of ViuHs are
covalently coupled to one another by a linker, the linker comprising one or more amino acids.
8. The polypeptide of any one of claims 1 to 7, wherein the variable region fragment
of the heavy chain antibody comprises an amino acid sequence at least 80%, 90%, 95%, 97%,
98%, 99%, or 100% identical to any one of SEQ ID Nos: 1 to 6 or 29 to 150.
9. The polypeptide of any one of claims 1 to 7, wherein the variable region fragment
of the heavy chain antibody comprises a complementarity determining region 1 (CDR1) as set
forth in any one of SEQ ID Nos: 7 to 12 or 151 to 272, a complementarity determining region 2
(CDR2) as set forth in any one of SEQ ID Nos: 13 to 18 or 273 to 394, and a complementarity
determining region 3 (CDR3) as set forth in any one of SEQ ID Nos: 19 to 24 or 395 to 516.
10 The polypeptide of any one of claims 1 to 7, wherein the amino acid sequence of
the VyH comprises:
(a) a CDRI sequence set forth in SEQ ID No: 7, a CDR2 sequence set forth
in SEQ ID No: 13, and a CDR3 sequence set forth in SEQ ID No: 19.
(b) a CDRI sequence set forth in SEQ ID No: 8, a CDR2 sequence set forth
in SEQ ID No: 14, and a CDR3 sequence set forth in SEQ ID No: 20
(c) a CDRI sequence set forth in SEQ ID No: 10, a CDR2 sequence set forth
WO 2020/008254 . PCT/IB2019/000687
(d) a CDRI sequence set forth in SEQ ID No: 11, a CDR2 sequence set forth
(e) a CDRI sequence set forth in SEQ ID No: 175, a CDR2 sequence set
forth in SEQ ID No: 297, and a CDR3 sequence set forth in SEQ ID No: 419.
[63) a CDRI sequence set forth in SEQ ID No: 218, a CDR2 sequence set
(g a CDRI sequence set forth in SEQ ID No: 219, a CDR2 sequence set
forth in SEQ ID No: 341, and a CDR3 sequence set forth in SEQ ID No: 463
(h) a CDRI sequence set forth in SEQ ID No: 243, a CDR2 sequence set
11 A polypeptide complex, wherein the polypeptide comprises a first component
polypeptide and a second component polypeptide, wherein the first component polypeptide and
the second component polypeptide are not covalently linked together and are coupled together
by a protein-protein interaction, a small molecule-protein interaction, or a small molecule-small
molecule interaction, wherein each of the first and the second component polypeptides comprise
a VzH which specifically binds an aquaculture-associated pathogen.
12 The polypeptide of any one of claims 1 to 10 or the polypeptide complex of claim
11, wherein the aquaculture-associated pathogen is a shellfish-associated bacterium
11, wherein the aquaculture-associated bacteria comprises a species of Vibrio
14. The polypeptide of any one of claims 1 to 10 or the polypeptide complex of claim
11, wherein the species of Vibrio is selected from the list consisting of V. adaptatus, V.
aerogenes, V. aestivus, V. aestuarianus, V. agarivorans, V. albensis, V. alfacsensis, V.
alginolyticus, V. anguillarum, V. areninigrae, V. artabrorum, V. atlanticus, V. atypicus, V.
zureus, V. brasiliensis, V. bubulus, V. calviensis, V. campbellii, V. casei, V. chagasii, V.
cholerae, V. cincinnatiensis, V. coralliilyticus, V. crassostreae, V. cyclitrophicus, V. diabolicus,
V. diazotrophicus, V. ezurae, V. fluvialis, V. fortis, V. furnissii, V. gallicus, V. gazogenes, V.
gigantis, V. halioticoli, V. harveyi, V. hepatarius, V. hippocampi, V. hispanicus, V. ichthyoenteri,

V.indicus, V. kanaloae, V. lentus, V. litoralis, V. logei, V. mediterranei, V. metschnikovii, V.
mimicus, V. mytili, V. natriegens, V. navarrensis, V. neonatus, V. neptunius, V. nereis, V.
nigripulchritudo, V. ordalii, V. orientalis, V. pacinii, V. parahaemolyticus, V. pectenicida, V.
penaeicida, V. pomeroyi, V. ponticus, V. proteolyticus, V. rotiferianus, V. ruber, V. rumoiensis,
V. salmonicida, V. scophthalmi, V. splendidus, V. superstes, V. tapetis, V. tasmaniensis, V.
tubiashii, V. vulnificus, V. wodanis, V. xuii, V. fischer, or V. hollisae

WO 2020/008254 » PCT/IB2019/000687
15 The polypeptide or the polypeptide complex of claim 14, wherein the VyH
specifically binds a Vibrio virulence factor
16 The polypeptide or the polypeptide complex of claim 14, wherein the VyH
specifically binds an antigen or polypeptide at least 60%, 70%, 80%, 90%, 95%, 98%, 99%, or
100% identical to SEQ IDs Nos: 25 or 26 or combinations thereof.
17. The polypeptide or the polypeptide complex of claim 15, wherein the Vibrio
virulence factor is PirA polypeptide, PirA-like toxin polypeptide, PirB polypeptide, or PirB-like
toxin.
11, wherein the aquaculture-associated pathogen is White Spot Syndrome Virus.
11, wherein the VyH specifically binds an antigen or polypeptide at least 60%, 70%, 80%, 90%,
95%, 98%, 99%, or 100% identical to SEQ IDs Nos: 27 or 28 or combinations thereof.
20. The polypeptide of any one of claims 1 to 10 or the polypeptide complex of claim
11, wherein the VyH specifically binds a polypeptide from VP24 or VP28, and combinations
thereof.
11, wherein the VyH can be pulled-down in a protein-protein binding assay by any of SEQ ID
Nos: 25 or 26 or 27 or 28
11, wherein the VyH survives in shrimp midgut extract for at least 1 minute, 5 minutes, 15
minutes, 30 minutes, 1 hour, 2 hours, 4 hours, or 24 hours.
23. A nucleic acid encoding the polypeptide of any one of claims 1 to 10 or 12 to 22

or the polypeptide complex of any one of claims 11 to 22.
24 A plurality of nucleic acids encoding the polypeptide complex of any one of
claims 11 to 22.
25 A cell comprising the nucleic acid of claim 23 or the plurality of nucleic acids of
claim 24.
26. The cell of claim 25, wherein the cell is a yeast cell
27. The cell of claim 26, wherein the yeast is of the genus Pichia.
28. The cell of claim 26, wherein the yeast is of the genus Saccharomyces.
29. The cell of claim 25, wherein the cell is a bacterial cell.
30 The cell of claim 29, wherein the bacteria is of the genus Escherichia.
31 The cell of claim 29, wherein the bacteria is a probiotic bacterium.
WO 2020/008254 w PCT/IB2019/000687
32 The cell of claim 31, wherein the probiotic bacteria is selected from the group
consisting of the genus Bacillus, the genus Lactobacillus, the genus Bifidobacteriunt.
33 The polypeptide of any one of claims 1 to 10 or 12 to 22 or the polypeptide
complex of any one of claims 11 to 22 further comprising a vitamin, an antibiotic, a hormone, an
antimicrobial peptide, a steroid, a probiotic, a probiotic, a bacteriophage, chitin, chitosan, B-1,3-
glucan, vegetable extracts, peptone, shrimp meal, krill, algae, B-cyclodextran, alginate, gum,
tragacanth, pectin, gelatin, an additive spray, a toxin binder, a short chain fatty acid, a medium
chain fatty acid, yeast, a yeast extract, sugar, a digestive enzyme, a digestive compound, an
essential mineral, an essential salt, or fibre.
34 A method of producing the polypeptide of any one of claims 1 to 10 or 12 to 22
or the polypeptide complex of any one of claims 11 to 22, comprising (a) incubating a cell of
any one of claims 25 to 32 in a medium suitable for secretion of the polypeptide from the cell;
and (b) purifying the polypeptide from the medium.
35. The polypeptide of any one of claims 1 to 10 or 12 to 22 or the polypeptide
complex of any one of claims 11 to 22 for use in reducing or preventing a fish or shellfish-
associated bacterial or viral infection in a human individual or a fish or shellfish
36 The polypeptide of any one of claims 1 to 10 or 12 to 22 or the polypeptide
complex of any one of claims 11 to 22 for use in reducing transmission or preventing
transmission of a fish or shellfish-associated bacterial or viral infection from a fish or shellfish to
another fish or shellfish or a human individual
37. The use of claim 35 or 36, wherein the shellfish is a species of crustaceans,
bivalvia, gastropods, cephalopods, octopus, squid, cuttlefish, clams, oysters, mussels, scallops,
cockles, whelks, winkles, shrimp, prawns, crawfish, crayfish, lobster, crabs, krill and barnacles.
38. The use of claim 35 or 36, wherein the fish is a species of catfish, carp, tilapia,
salmon, tuna, eel, milkfish, trout, snakehead, loach, and perch.
39 The use of claim 35 or 36 wherein the polypeptide is adapted for introduction to

the alimentary canal orally or rectally, provided to the exterior surface, provided to the medium
in which the animal dwells, provided by injection, provided intravenously, provided via the
respiratory system, provided via diffusion, provided via absorption by the endothelium or
epithelium, or provided via a secondary organism such as a yeast, bacterium, algae,
bacteriophages, plants and insects to a host.
40. A method of reducing transmission or preventing transmission of a fish or
shellfish-associated bacterial or viral infection from a fish or shellfish to another fish or shellfish
comprising administering the polypeptide of any one of claims 1 to 10 or 12 to 22 or the
WO 2020/008254 PCT/1B2019/000687
41
polypeptide complex of any one of claims 11 to 22 to an aquaculture comprising fish or

shellfish
WO 2020/008254 PCT/1B2019/000687
17
o2
|
|
ciz
FIG. 1B
VHH
Amino Acid Sequence
FR1 ] cori]FR2[cor2] FR3 | CDR3 [FR4}-C

1
2
2 s
0 i
PCT/1B2019/000687
i
Antigen-Coated
FES-Coated
e
ey
E——
e
Antigen-Coated
2 s, i
a
S
§ e—
i
s
NBX Number
NBX Number
@ I sz
FIG.2B
FIG.2A
s
* — s
2/7
i
s
O
it
P
— i
e o - @ i
£=1 (=3 f=1 £=3
s
oM pereo) gad Sy s
MM paeo) uabguy Sy s
2150
WO 2020/008254
i
2 < o
= S =
0.8
10
13 PoIR0D S8d STy
MM paIEo] usBiuy SOy
WO 2020/008254 PCT/1B2019/000687
317
B
e Antigen-Coated
== PBS-Coated
4. Upper Limit of Detec s S
g T oad
R
33
§ L
w0
2 8 2
34
<
8
14
J bR LE N . A " N . A
SEESSSES
FPEIPIIFSEFFPLEL
PP PP
S SSSSSSSSSSS
NBX Number
WO 2020/008254 PCT/1B2019/000687
47
FIG. 2D
- Antigen-Coated
trd
W PRS-Costed
Agen Antigen Coated Wellf
Bg5; PBS Coated Well
L
N STEIR
LR IL s i<kks ‘ B bbb§
LEER L. N
SRR
RO
HBX Number
PCT/1B2019/000687
k=l Wﬁ
m d - s &v\v
Q% i B
Q8 Am:*.@v
5 O K
2 0 e RN
NEX Number
NBX Number
&g G
-" KA
L %9,
FIG. 2E
FIG. 2F
&.r
S
517
gp@@,@
Aantigen-Coated
%%,
PES-Coated
o
& » ~
—
- P &m.%
1,
119 PoIEOD SHd Py (4
e
B
HiBp payeoD uabyuy By
WO 2020/008254
8.5
284
- -
Hep pEIeDD S84 Py
o, payeoz uaBhuy %hy
SP8OXaN
XEN - ¢
QWL WPz HST Mz MZOF OB ST §
B daiys iBpiu duiniys
QZ80XEN SZO0XaN
¥ "Old
International application No.
INTERNATIONAL SEARCH REPORT
PCT/IB2019/000687
A, CLASSIFICATION OF SUBJECT MATTER
IPC: CO7K 16/12 2006.01), AGIK 39/42 (2006.01), AGIP31/12(200601), CO7K 16/00(2006.01),
CO7K 16/08 (2006.01), CI2N 1/19 (2006.01) (more IPCs on the last page)
According to International Patent Classification (IPC) or to both national classification and IPC
B. FIELDS SEARCHED
Minimum documentation scarched (classification system followed by classification symbols)

IPC: COTK 16/12 (2006.01), A61K 39/42 (2006.01), A61P 31/12 (2006.01), COTK 16/00 (2006.01) , COTK 16/08 (2006.01) ,
CI2N 1/19 (2006.01) , CI2N 1/21(2006.01), CI2N 15/13 (2006.01) , C12P 21/00 (2006.01),, C40B 40/08 (2006.01)
Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched
Electronic database(s) consulted during the intemational scarch (name of database(s) and, where practicable, search terms used)
[Databases: CIPO library discovery tool, Scopus, PubmedCentral, Questel-Orbit, GQPAT, GeneSeq Protein, Uniprot, RefSeq, GenPept, IPI,
IGBLAST Protein, PDB protein, ENSEMBL.
[Keywords: heavy chain antibody, Vhh, nanobody, vibrio, aquaculture associated pathogen, whitc spot syndrome virus.
SEQ ID NO: 1-6 and 29-150 @80% identity, 7-24 and 151-516 @100% identity.
C. DOCUMENTS CONSIDERED TO BE RELEVANT
Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.
EBRAHIMIZADEH W et al. “Isolation and characterization of protective anti-LPS nanobody

X against V. cholera O1 recognizing Inaba and Ogawa serotypes” Appl Microbiol Biotechnol. 2013 1,2,47, 11-15
and 2240
May; 97(10):2257-66. doi: 10.1007/500253-012-4518-x.
*the whole document®
Panning anti-LPS nanobody as a capture target to enrich Vibrio fluvialis” Biochem Biophys Res
P,X | Commun.2019 May 7: 512(3):531-536. doi: 10.1016/j.bbrc.2019.03.104. Epub 2019 Mar 22. 1-40
*the whole document®
A WO 2017/199094 A1, ABNOUSI H, 23 November 2017 (23-11-2017). 1-40

*the whole document*
[V Further documents are listed in the continuation of Box C. ¥ Sce patent family annex.
+[special categories of cited documents: “1 [later document published after the intemational filing date or priority
“A” |document defining the general state of the art which is not considered date and not in conflict with the application but cited to understand
to be of particular relevance the principle or theory underlying the invention
“D” |document cited by the applicant in the international application “X” [document of particular relevance; the claimed invention cannot be
“E |earlier application or patent but published on or after the international considered novel or cannot be considered to involve an inventive
filing date step when the document is taken alone
“L” |document which may throw doubts on priority claim(s) or which is “Y” [document of particular relevance; the claimed invention cannot be
cited to establish the publication date of another citation or other considered to involve an inventive step when the document is
special reason (as specified) combined with one or more other such documents, such combination
0" |document referring to an oral disclosure, use, exhibition or other means being obvious to a person skilled in the art
& |document member of the same patent family
“P |document published prior to the international filing date but later than
b date claimod
Date of the actual completion of the international search Date of mailing of the international scarch report
12 December 2019 (12-12-2019) 12 December 2019 (12-12-2019)
Name and mailing address of the ISA/CA Authorized officer

Canadian Intellectual Property Office
Place du Portage I, C114 - 1t Floor, Box PCT Mostapha Bayaa (819) 639-7743
50 Victoria Street
Gatincau, Quebec K1A 0C9
Facsimile No.: 819-053-2476
Form PCT/ISA/210 (second sheet ) (July 2019) Page 3 0f 6
INTERNATIONAL SEARCH REPORT PCT/IB2019/000687
C (Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT
Category* | Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.
A WO 2015/145250 A2, BANERJEE SK et al. 1 October 2015 (01-10-2015). 1-40

*the whole document*
Form PCT/ISA/210 (continuation of second sheet) (July 2019) Page 4ol 6

INTERNATIONAL SEARCH REPORT
PCT/IB2019/000687
Box No. 1 Nucleotide and/or amino acid sequence(s) (Continuation of item 1.c of the first sheet)
1. With regard to any nucleotide and/or amino acid sequence disclosed in the interational application, the international search was carried out
on the basis of a sequence listing:
a [ forming part of the international application as filed:
I in the form ofan Annex C/ST.25 text file.
™' on paper or in the form of an image file.
b. [ furnished together with the international application under PCT Rule 13fer.1(a) for the purposes of intemational scarch only in the
form ofan Annex C/ST.25 text file.
¢. ¥ furnished subsequent to the international filing date for the purposes of international search only:
[¥in the form of an Annex C/ST.25 text file (Rule 137er.1(a)).
I™ on paper or in the form of an image file (Rule 13fer.1(b) and Administrative Instructions, Section 713).
2. I Tnaddition, in the case that more than one version or copy of a sequence listing has been filed or furnished, the required statements that
the information in the subsequent or additional copies is identical to that in the application as filed or does not go beyond the application
as filled, as appropriate, were furnished.
3.Additional comments:
Form PCT/ISA/210 (continuation of first sheet (1)) (July 2019) Page 2016
INTERNATIONAL SEARCH REPORT PCT/IB2019/000687
CI2N 1/21 (2006.01), CI2N 15/13 (2006.01), CI2P 21/00 (2006.01), C40B 40/08 (2006.01)
Form PCT/ISA210 (extra sheet) (July 2019) Page 6 0f 6

INTERNATIONAL SEARCH REPORT International application No.
Information on patent family members PCT/IB2019/000687
Patent Document Publication Patent Family Publication

Cited in Scarch Report ~ Date Member(s) Date
WO2017199094A1 23 November 2017 (23-11-2017) US2019202896A1 04 July 2019 (04-07-2019)
WO2015145250A2 01 October 2015 (01-10-2015) WO2015145250A3 21 January 2016 (21-01-2016)
AU2015237933A1 20 October 2016 (20-10-2016)
BR112016022375A2 15 January 2019 (15-01-2019)
CA2943903A1 01 October 2015 (01-10-2015)
CN107074934A 18 August 2017 (18-08-2017)
EP3122773A2 01 February 2017 (01-02-2017)
IN1100MU2014A 02 October 2015 (02-10-2015)
MX2016012597A 27 April 2017 (27-04-2017)
US2017152305A1 01 June 2017 (01-06-2017)
Form PCT/ISA/210 (patent family annex ) (July 2019) Page 50l 6

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(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)

Floor, Vancouver, BC V6E 0C3 (CA). VAN PETEGEM,

[Continued on next page]

ANTIBODIES AGAINST AQUACULTURE DISEASE-CAUSING AGENTS AND USES

CROSS-REFERENCE TO RELATED APPLICATIONS

FIELD OF THE INVENTION

BACKGROUND OF THE INVENTION

SUMMARY OF THE INVENTION

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] As referred to herein, “shellfish” refers to any aquatic exoskeleton-bearing invertebrate

crustaceans, bivalvia, gastropods, cephalopods, octopus, squid, cuttlefish, clams, oysters,

limitation, to Vibrio species, Aeromonas species, Edwarsiella species, Streptococcus species,

constitute specific binding.

methods described herein do not utilize conventional antibodies

supplement, a premix, a medicine, a therapeutic, a drug, a diagnostic tool, a component or

DETAILED DESCRIPTION OF THE INVENTION

a dromedary, camel, llama, alpaca, vicuna or guanaco, without limitation. In certain

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

NBX Amino acid sequence Antigen

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

NBX Amino acid sequence Antigen

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 1 Unique SEQ IDs for VyH antibodies of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Table 2 Unique SEQ IDs for VyH CDRs of this disclosure

Antibodies recombinantly expressed

limited to these forms.

peptide, a steroid, a probiotic, a bacteriophage, chitin, chitosan, B-1,3-glucan, vegetable extracts,

anguillarum, V. areninigrae, V. artabrorum, V. atlanticus, V. atypicus, V. azureus, V.

brasiliensis, V. bubulus, V. calviensis, V. campbellii, V. casei, V. chagasii, V. cholerae, V.

cincinnatiensis, V. coralliilyticus, V. crassostreae, V. cyclitrophicus, V. diabolicus, V.

diazotrophicus, V. ezurae, V. fluvialis, V. fortis, V. furnissii, V. gallicus, V. gazogenes, V.

gigantis, V. halioticoli, V. harveyi, V. hepatarius, V. hippocampi, V. hispanicus, V. ichthyoenteri,

mimicus, V. mytili, V. natriegens, V. navarrensis, V. neonatus, V. neptunius, V. nereis, V.

nigripulchritudo, V. ordalii, V. orientalis, V. pacinii, V. parahaemolyticus, V. pectenicida, V.

penaeicida, V. pomeroyi, V. ponticus, V. proteolyticus, V. rotiferianus, V. ruber, V. rumoiensis,

V. salmonicida, V. scophthalmi, V. splendidus, V. superstes, V. tapetis, V. tasmaniensis, V.

tubiashii, V. vulnificus, V. wodanis, V. xuii, V. fischer, or V. hollisae

parahaemolyticus OX=670 GN=vp19 PE=4 SV=1

SWCTVHGFEHMLIW QKIKELKKVDVFVHSNLISY SPAVGFPSGNFNYIATGTEDEIPQPL

>tr/QOE7K6|Q9E7K6_WSSV Major structural protein VP24 OS=White spot syndrome virus

>tr|A6ZI33|A6Z133_WSSV Coat protein OS=White spot syndrome virus 0X=92652 GN=VP28

2. Production of NBXs and panning

NBX0725, NBX0730, NBX0738, NBX0739, NBX0745, and NBX0746.

Purification of VyHs from E. coli

[0060] The following references are incorporated by reference in their entirety

Hor, L. I, Lo, C. F. (2015) The opportunistic marine pathogen Vibrio parahaemolyticus

4. FAO Fisheries and Aquaculture (2013) Report of the FAO/MARD Technical

Lightner, D. V. (2013) Determination of the infectious nature of the agent of acute

M., El-Ashram, A. M. M. (2018) Molecular characterization, antibiotic resistance pattern and