REVIEW REVIEW

mAbs 2:4, 365-378; July/August 2010; © 2010 Landes Bioscience

Mining human antibody repertoires

Roger R. Beerli1,† and Christoph Rader2
Cytos Biotechnology AG; Schlieren, Switzerland; 2Experimental Transplantation and Immunology Branch; Center for Cancer Research; National Cancer Institute;
1

National Institutes of Health; Bethesda, MD USA

†
Current Address: Intercell AG; Wagistrasse 25; CH-8952 Schlieren, Switzerland

Key words: human monoclonal antibodies, B cells, hybridoma technology, display technologies, antibody libraries,
antibody engineering

Abbreviations: Ig, immunoglobulin; mAbs, monoclonal antibodies; HAMA, human anti-mouse antibody; HARA, human anti-rat
antibody; HACA, human anti-chimeric antibody; HAHA, human anti-human antibody; FDA, Food and Drug Administration;
EMA, European Medicines Agency; GC, germinal centers; EBV, Epstein-Barr virus; AID, activation-induced deaminase;
PCR, polymerase chain reaction; RT, reverse transcriptase; CDR, complementarity-determining region; FACS,
fluorescence-activated cell sorting; GMP; good manufacturing practice; pAbs, polyclonal antibodies

Human monoclonal antibodies (mAbs) have become drugs of
choice for the management of an increasing number of human
diseases. Human antibody repertoires provide a rich source for
human mAbs. Here we review the characteristics of natural and
non-natural human antibody repertoires and their mining with
non-combinatorial and combinatorial strategies. In particular,
we discuss the selection of human mAbs from naïve, immune,
transgenic and synthetic human antibody repertoires using
methods based on hybridoma technology, clonal expansion
of peripheral B cells, single-cell PCR, phage display, yeast
display and mammalian cell display. Our reliance on different
strategies is shifting as we gain experience and refine methods
to the efficient generation of human mAbs with superior
pharmacokinetic and pharmacodynamic properties.
Introduction
Human antibody repertoires are collections of human immuno-
globulin (Ig) genes that encode human heavy and light chains.
These include V H, D, JH and CH gene segments of the heavy
chain locus on chromosome 14; Vk, Jk and C k gene segments of
the kappa light chain locus on chromosome 2; and Vl, Jl and Cl
gene segments of the lambda light chain locus on chromosome
22 in the human genome. In addition, transgenic animals with
complete or partial human antibody repertoires have been gener-
ated. Human antibody repertoires can be mined in vivo and in
vitro for human monoclonal antibodies (mAbs).
Chimeric, humanized and human mAbs, which collectively
are the prevailing formats of therapeutic mAbs, share human
constant domains but are discerned by the origin of their variable
Correspondence to: Roger R. Beerli and Christoph Rader;
Email: rbeerli@intercell.com and raderc@mail.nih.gov
Submitted: 03/26/10; Accepted: 04/27/10
Previously published online:
www.landesbioscience.com/journals/mabs/article/12187
domains. Human mAbs (also referred to as fully human mAbs)
are defined as having variable domains that are entirely derived
from human antibody repertoires, whereas chimeric and human-
ized mAbs feature variable domains that originate from non-
human antibody repertoires. A key feature of human mAbs is
their ability to fully blend in with the human immune system.
Being indistinguishable from endogenous human antibodies,
human mAbs are generally anticipated to have superior pharma-
cokinetic and pharmacodynamic properties compared to mAbs
from nonhuman antibody repertoires. Ideally, human mAbs
are not recognized as foreign by the human immune system,
i.e., are not immunogenic. Human and humanized mAbs have
been found to be less immunogenic than nonhuman and chi-
meric mAbs, the latter of which trigger human anti-mouse anti-
body (HAMA), human anti-rat antibody (HARA) and human
anti-chimeric antibody (HACA) responses.1 It is expected, but
remains to be substantiated, that human mAbs are generally less
immunogenic than humanized mAbs.2,3 Human anti-human
antibody (HAHA) responses are mainly due to idiotypic, allo-
typic and glycosylation differences between human mAbs and
endogenous human antibodies. Although potentially beneficial
for certain therapeutic applications,4 it is likely that fine tuning
of human mAb engineering and manufacturing will reduce the
remaining immunogenicity of human mAbs.
Hybridoma technology 5 has been extremely successful in the
generation of mouse and rat mAbs; however, the initial failure
to robustly apply this method to the generation of human mAbs
paired with the prospect of lucrative intellectual property accel-
erated the development of a variety of alternative methods over
the past three decades.2,6-8 The medical and commercial success
of mAbs in the management of human disease drove this growth
and diversification.9,10 Although only six out of 28 mAbs that were
approved by the US Food and Drug Administration (FDA) and/
or the European Medicines Agency (EMA) are human mAbs
(Table 1), four of these antibodies were approved as recently
as 2009, and their proportion is likely to increase, owing to an
unremitting pipeline of human mAbs in Phase 3 clinical trials.11

www.landesbioscience.com mAbs 365

 Table 1. Human mAbs approved in the United States and the European Union
Generic name
Format (origin)
(trade name; company)
Adalimumab
IgG1k (phage display)
(Humira®; Abbott)
Panitumumab
IgG2k (transgenic mouse)
(Vectibix®; Amgen)
Golimumab
IgG1k (transgenic mouse)
(Simponi®; Johnson & Johnson)
Canakinumab
IgG1k (transgenic mouse)
(Ilaris®; Novartis)
Ustekinumab
IgG1k (transgenic mouse)
(Stelara®; Johnson & Johnson) ­(common p40 subunit)
Ofatumumab
IgG1k (transgenic mouse)
(Arzerra®; Genmab)
Here we review the generation of human mAbs from naïve,
immune, transgenic and synthetic human antibody repertoires.
Strictly speaking, only human mAbs from naïve and immune
repertoires, which we summarize as natural repertoires in the
next section, as well as human mAbs from transgenic reper-
toires, which nonetheless are products of artificial immune sys-
tems, meet the above stated criterion of having variable domains
that are entirely derived from human antibody repertoires and
are therefore indistinguishable from endogenous human anti-
bodies. By contrast, human mAbs from synthetic repertoires
generally contain artificial sequences that are not encoded in
the human genome and cannot be generated by natural pro-
cesses such as V HDJH, VkJk and VlJl rearrangements and
somatic hypermutation. Transgenic and synthetic repertoires
are summarized as non-natural repertoires in a subsequent sec-
tion. Furthermore, we distinguish between non-combinatorial
and combinatorial strategies for mining human antibody rep-
ertoires. Non-combinatorial strategies retrieve human mAbs
from single B-cells with the original heavy and light chain
pairs (Fig. 1). Combinatorial strategies involve human anti-
body libraries with randomly combined heavy and light chains
(Fig. 2). Hybridoma technology and phage display technology
exemplify non-combinatorial and combinatorial mining tools,
respectively. Both approaches have led to FDA- and EMA-
approved human mAbs (Table 1).
Natural Repertoires
Each B cell encodes a single antibody with defined specificity.
The clonal diversity of the natural human antibody repertoire
is defined by the number of B cells that encode antibodies with
unique specificities, i.e., the number of independent clones. In
an individual, this number is estimated to be at least 108. B
cells from primary and secondary lymphoid tissues along with
B cells in circulation, i.e., in peripheral blood, constitute the
natural human antibody repertoire. The natural repertoire
can be divided into naïve and immune repertoires depending
on whether or not it has been shaped by antigen encounter.
The immune repertoire has been extensively mined for human
366
Antigen Indications Approval
Rheumatoid arthritis; juvenile idiopathic
2002 (FDA),
TNFa arthritis; psoriatic arthritis; plaque psoriasis;
2003 (EMA)
ankylosing spondylitis; Crohn’s disease
2006 (FDA),
EGFR Metastatic colorectal cancer
2007 (EMA)
Rheumatoid arthritis; psoriatic arthritis; 2009 (FDA),
TNFa
ankylosing spondylitis 2009 (EMA)
Cryopyrin-associated periodic syndrome 2009 (FDA),
IL-1b
(CAPS) 2009 (EMA)
IL-12 and IL-23 2009 (FDA),
Plaque psoriasis
2009 (EMA)
2009 (FDA),
CD20 Chronic lymphocytic leukemia
2010 (EMA)
mAbs, first with non-combinatorial and later with combina-
torial strategies. By contrast, combinatorial strategies have
dominated access to human mAbs from the naïve repertoire
(Table 2).
The bone marrow is a primary lymphoid tissue of interest
for mining natural human antibody repertoires. The principle
site of differentiation of hematopoietic stem cells into immature
B cells,12 the bone marrow generates pro-B cells and pre-B cells
with partially rearranged heavy and light chain genes. While
these partially developed human antibody repertoires are gener-
ally not suitable for mining human mAbs, surrobody libraries
that pair rearranged heavy chains with surrogate light chains of
pre-B cells have been described recently.13 The ultimate products
of B-cell development in the bone marrow are immature B cells
that express cell surface IgM. Upon exiting the bone marrow and
entering the circulation, immature B cells mature to naïve B cells,
the latter of which express cell surface IgD in addition to IgM.
Immature and naïve B cells collectively provide a fully developed
human antibody repertoire that has not been shaped, at least not
extensively, by antigen encounter and is therefore termed primary
repertoire or naïve repertoire.
When naïve B cells encounter new antigens that bind to
their surface Ig they become activated B cells, proliferate and go
through differentiation processes that typically involve somatic
hypermutation and class switch recombination from the earliest
expressed IgM isotype to IgG, IgE and IgA isotypes. These pro-
cesses are T-cell-dependent and take place in germinal centers
(GC) of secondary lymphoid tissues such as lymph nodes, spleen
and tonsils. Post-GC B cells with affinity and isotype matured
antibodies differentiate further into antibody secreting cells and
memory B cells.14 Antibody secreting cells can be divided into
proliferating plasma blasts and fully differentiated plasma cells
that no longer proliferate. Plasma cells exit secondary lymphoid
tissues, enter the circulation, and migrate to the bone marrow
where they may survive for weeks, months, or even years in spe-
cialized niches. Antibody titers in circulation are maintained by
plasma cells residing in the bone marrow.15 In contrast to anti-
body secreting cells, memory B cells do not secrete antibodies but
express cell surface Ig. Memory B cells are found in secondary
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 Figure 1. Non-combinatorial mining of human antibody repertoires. (A) Human antibody repertoires consist of polyclonal mixtures of a variety of B
cells that express functional human antibodies, including naive and post-GC B cells that represent naïve and immune repertoires, respectively.
Post-GC B cells, which differentiate into antibody secreting cells and memory B cells, provide a human antibody repertoire that has been extensively
mined with non-combinatorial strategies. (B) Hybridoma technology (top), EBV immortalization (center), and single-cell sorting (bottom) allow the
isolation and expansion of monoclonal B cells which are then screened with an antigen of interest. Cell surface Ig can be utilized to first enrich B cells
that express human antibodies to the antigen of interest. (C) Heavy and light chain cDNAs are amplified from mRNA of monoclonal B cells and B-cell
lines by RT-PCR. (D) After cloning heavy and light chain cDNAs into ectopic expression vectors, human mAbs are purified from the supernatant of
transfected mammalian cells (typically Chinese hamster ovary CHO or mouse myeloma NS0 cells) in high yields.

lymphoid tissues, in circulation and in the bone marrow. Upon
recall antigen encounters, memory B cells rapidly differentiate
into antibody secreting cells. Both memory B cells and plasma
cells can be long-lived, maintaining immunological memory for
a lifetime.16
The secondary repertoire, or immune repertoire, is largely
defined by the pool of post-GC B cells, plasma blasts, plasma
cells and memory B cells. Expression of CD27 distinguishes
this B-cell pool from other B cells. Antibody-secreting cells
and memory B cells can also arise from T-cell-independent
processes that play a role in both adaptive and innate immunity.
Adaptive immunity to T-cell-independent antigens such as
carbohydrates and lipids contributes to the immune repertoire.
By contrast, cells expressing natural antibodies, which play a
key role in innate immunity,17 can be considered to belong to
the naïve repertoire. These two types are difficult to distinguish
and typically share IgM isotype, low affinity and polyreactivity.
They illustrate that immune and naïve repertoires often
overlap without clear anatomical localizations or functional
distinctions.
Human immune repertoires are highly attractive because
they contain antibodies of high affinity with minimal immu-
nogenicity and off-target reactivity. Unlike nonhuman immune
repertoires that have been mined extensively for mAbs, the
human immune repertoire lacks both availability and accessi-
bility. First of all, humans cannot be exposed to antigens at
will, limiting available immune repertoires to those induced
by infections, vaccinations, autoimmunity and alloimmunity.
Furthermore, secondary lymphoid tissues such as the spleen,
which is a preferred source of mouse, rat and rabbit immune
repertoires for mAb mining, are difficult to access in humans.
Although tonsillectomy provides access to tonsils, it generally
does not coincide with an immune response of interest. Access
to the human immune repertoire is therefore generally limited
to peripheral blood and, to a lesser extent, bone marrow. As dis-
cussed above, these compartments also contain the naïve rep-
ertoire. Combinatorial strategies allow the retrieval of human
mAbs from the naïve repertoire. Typically, these mAbs need
to be improved by affinity maturation in vitro to compensate
for their lack of efficient selection by antigens in vivo. The next

www.landesbioscience.com mAbs 367

 Figure 2. Combinatorial mining of human antibody repertoires. Polyclonal or oligoclonal mixtures of human B cells (A) are used in bulk to isolate
mRNA and amplify cDNA of heavy and light chains (B). (C) Heavy and light chain cDNAs are cloned into display vectors that afford antibody libraries
with a physical linkage of genotype (cDNA) and phenotype (protein). Shown as examples are filamentous phage that encode and display Fab (top) and
virus-infected mammalian cells that express and display scFv (bottom). Note that heavy and light chains are randomly combined. (D) Antibody
libraries are then selected by several rounds of panning on immobilized antigen (top) or screened by a single round of FACS using fluorescently
labeled antigen (bottom). (See text for details). The physical linkage of genotype and phenotype allows the enrichment and amplification of displayed
antibodies. (E) Human mAbs from antibody libraries are manufactured the same way as human mAbs from monoclonal B cells and B-cell lines.

Table 2. Principal mining tools for human antibody repertoires

Mining tools
Repertoire
Non-combinatorial Combinatorial
Naïve Phage display; yeast display
Natural Hybridoma technology; clonal expansion; Phage display; yeast display; mammalian cell
Immune
single-cell PCR display
Transgenic Hybridoma technology
Non-natural
Synthetic Phage display; yeast display; ribosome display

two sections discuss various features of naïve and immune rep-
ertoires that are exploited in non-combinatorial and combinato-
rial strategies for mining human mAbs.
Non-Combinatorial Mining
Hybridoma technology. The advent of hybridoma technology had
a tremendous impact on biomedical research, yielding mAbs that
have become an indispensable part of everyday laboratory work.
Today rodent mAbs against a plethora of antigens are widely
available and can be produced in almost unlimited quantities due
to this robust underlying technology.5 Unfortunately, the therapeu-
tic use of rodent mAbs in humans is limited by their propensity to
induce strong HAMA and HARA responses that quickly neutral-
ize the rodent mAb. Given this, the mining of human antibody
repertoires using hybridoma technology was investigated exten-
sively. Despite significant efforts, the adaptation of hybridoma
technology to the generation of human mAbs has been a slow and
difficult process due to the limited number of B cells stimulated
with an antigen of interest and a lack of suitable drug-resistant

368 mAbs Volume 2 Issue 4

myeloma cell fusion partners for the immortalization of human
B cells. Thus, mouse myeloma cells were initially used for the
generation of mouse-human heterohybridomas.18-20 Such hybrid
cells were shown to have a strong tendency to lose human chro-
mosomes, including chromosome 2 carrying the kappa light chain
locus.21 While this precluded an extensive use of mouse myeloma
cells for human mAb production, protocols have been established
that may ameliorate the undesirable behavior of heterohybridomas
and allow long-term production of human mAbs.22
Given the genetic instability of heterohybridomas, the use of
alternative fusion partners, particularly human lymphoblastoid
cell lines and human myeloma cells, has been explored exten-
sively.23-28 While in principle successful, the yield of viable hybrid-
omas is typically too poor for practical applications. In addition,
such hybridomas often secrete permutated Igs derived from
both fusion partners. The use of certain mouse-human hybrid
cells including heteromyelomas and heterohybridomas as fusion
partner appears to resolve these problems, while at the same time
generating stable antibody producing hybridomas.29-32 In addi-
tion, a promising human myeloma cell line allowing efficient
generation of stable and highly productive human hybridomas
was described, even though a more widespread use of this cell line
awaits the production of a subline lacking expression of an endog-
enous l chain.33 Finally, the use of genetically engineered mouse
myeloma cells allows for highly efficient formation of stable het-
erohybridomas,34 with a propensity to produce affinity-matured
IgG antibodies when appropriately pre-treated CD27-positive B
cells are used for fusion.35
As mentioned above, human hybridoma technology not only
suffered from the lack of suitable fusion partners, but also from
a shortage of antigen-stimulated B cells. Because ethical consid-
erations typically prevent antigen exposure at will in humans,
alternative methods to improve access to the human immune
repertoire were developed. Human peripheral blood lymphocytes
transplanted into immune-compromised mice were shown to
give rise to a secondary humoral immune response upon antigen
stimulation,36,37 and provide access to the human memory com-
partment through hybridoma technology.38,39
In summary, each of the methods described is either labor-
intensive or suffers from drawbacks when compared to the classi-
cal hybridoma technology. Thus, hybridoma technology has not
become the method of choice for mining human antibody rep-
ertoires, leading to tremendous efforts to develop the alternative
methods discussed in the following sections.
Clonal expansion of peripheral B cells. One of the most
popular methods for harnessing the human immune response
involves immortalization of peripheral B cells with Epstein-Barr
virus (EBV). The first demonstration of EBV immortalization
of human B cells into lymphoblastoid cell lines in vitro, was
described almost 40 years ago.40 There have been numerous
reports since then, although initial success was limited due to low
efficiency of B-cell immortalization and cloning and the small
quantities of antibody secreted.41,42 Better cloning efficiency, sta-
bility and productivity can be achieved when EBV-immortalized
antibody secreting cells are fused with mouse myeloma or mouse-
human heteromyeloma cells.43,44 Thus, multi-step procedures
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involving EBV-immortalization of peripheral B cells followed by
fusion with myeloma cells and cloning of hybridomas were estab-
lished.45-48 This method has yielded several human mAbs with
therapeutic potential, including neutralizing antibodies against
the hemagglutinin protein derived from the 1918 H1N1 pan-
demic influenza virus.49
As an alternative to the formation of hybridomas, EBV-
transformed cells have been infected with a retrovirus encoding
the ras oncogene, yielding cells with similar cloning efficiencies
and levels of antibody secretion.50 The most significant improve-
ment of EBV immortalization came from the observation that
addition of the TLR9 ligand CpG to human B cells dramatically
increases the efficiency of EBV immortalization and facilitates the
establishment of clonal lines stably secreting specific antibody. In
this manner, mAbs potently neutralizing the SARS corona virus
or the H5N1 Influenza A virus were isolated.51,52
Strategies allowing the long-term growth and cloning of
human B cells, without immortalization by transformation with
a virus or an oncogene, have also been explored. In initial experi-
ments, short-term B-cell clones were formed when cultured in the
presence of EL-4B5 thymoma cells in conjunction with human
T-cell plus macrophage supernatant.53,54 Improved proliferation
and antibody production were found under more defined condi-
tions, when B cells were activated by IL-4 treatment and CD40
triggering.55,56 A combination of both methods together with
further refinements proved extremely efficient, allowing direct
establishment of viable B-cell cultures from single, antigen-
specific B cells.57,58 Similar to EBV immortalization, these B-cell
clones secrete sufficient amounts of antibody into the superna-
tant, and production of specific antibody can be validated prior
to cloning of the variable domains. Further, high throughput
screens based on short term clonal expansion of activated periph-
eral B cells are possible and have led to the identification of HIV-1
neutralizing antibodies.59
Recently, a promising method involving genetic reprogram-
ming of human memory B cells was described.60 Retrovirus-
mediated expression of Bcl-6 and Bcl-x L in peripheral memory B
cells, combined with culturing on CD40L-expressing L cells in
the presence of IL-21, converts them to highly proliferating, cell
surface Ig-positive, antibody secreting B cells that can be cloned
efficiently. The proteins Bcl-6 and Bcl-x L are typically expressed
in GC B cells, and the resulting cells have features of GC B cells,
including expression of activation-induced deaminase (AID). It
is of interest whether this method will find broader application in
the generation of human mAbs.
Single-cell PCR. The most straightforward, though tech-
nically challenging, non-combinatorial strategy for mining
human antibody repertoires is single-cell polymerase chain
reaction (PCR). If both heavy and light chain variable domains
can be amplified in an efficient and reliable manner from single
cells, then antibodies with original heavy and light chain pair-
ings can be cloned on a cell-by-cell basis without the need to
grow individual B-cell clones. In principle, either memory B
cells or plasma cells can serve as a source of rearranged anti-
gen-binding sites. As discussed, peripheral memory B cells are
known to persist for many years after antigen exposure, thereby
369
offering a diverse and attractive repertoire of antigen specifici-
ties readily available in the blood. As memory B cells contain
only relatively small quantities of specific RNA, the successful
cloning of both heavy and light chain variable domains from
a single B cell poses a significant technical challenge. In spite
of this, antibodies have successfully been cloned from single
peripheral B cells, including mAbs against self-antigens,61,62 tet-
anus toxoid57 and HIV-1 gp140.63 The efficiency of cloning rel-
evant antibodies has been improved, e.g., by enriching specific
B cells using antigen-coated magnetic beads,57 or by directly
isolating antigen-specific B cells by fluorescence-activated cell
sorting (FACS).63
Compared to memory B cells, plasma cells have the disad-
vantage that they are present in the blood only for short periods
of time after antigen exposure, after which they are only acces-
sible in the bone marrow. Nonetheless, a known antigen exposure
or challenge, e.g., a booster immunization, induces circulating
plasma cells, of which a majority is specific for the antigen after
approximately one week.64 Although no highly plasma cell-
specific marker is known to date, they are readily isolated by
FACS, e.g., based on their high expression of CD19 and CD38,
and intermediate expression of CD45.65 These cells represent
dedicated antibody factories and contain huge amounts of spe-
cific heavy and light chain mRNAs, thereby facilitating antibody
cloning by reverse transcriptase (RT)-PCR. Numerous human
mAbs have been cloned from plasma cells to date, including
mAbs against tetanus toxoid and influenza virus.64-66 Recently,
using a method related to microengraving,67 microwell array
chips were successfully used for high-throughput screening of
human antibodies secreted by single CD138 + plasma cells fol-
lowed by RT-PCR cloning.68
Combinatorial Mining
Combinatorial mining of human antibody repertoires is inter-
twined with the concept of antibody libraries.8,69 Antibody librar-
ies randomly combine heavy and light chain cDNAs obtained
through RT-PCR from mRNA of B-cell pools into expression
vectors that afford a compartmental or physical linkage of phe-
notype (protein) and genotype (cDNA). In contrast to non-
combinatorial mining, the source of human antibody repertoires
subjected to combinatorial mining is B-cell mRNA rather than B
cells. Thus, antibody secreting cells, due to their higher heavy and
light chain gene transcript levels compared to other B cells, can
have much greater representation in antibody libraries. This bias
can be highly desirable for the combinatorial mining of immune,
but not naïve, repertoires. To increase the diversity of indepen-
dent clones in antibody libraries from naïve repertoires, RT-PCR
of the heavy chain mRNA can be restricted to the IgM isotype.
However, combinatorial strategies have inherent biases due to the
RT-PCR cloning of heavy and light chain cDNAs and due to
the expression and folding of non-natural antibody fragments in
non-B-cell environments. Nonetheless, high-throughput DNA
sequencing recently confirmed that large antibody libraries can
accurately mimic natural human antibody repertoires in terms of
molecular diversity.70,71
370 mAbs
Like other cDNA libraries, the first antibody libraries were
based on lambda phage vectors that carried heavy and light
chain cDNAs and facilitated their expression upon infection of
Escherichia coli bacteria.72 Lysis plaques of lambda phage on bac-
terial lawns were then screened with antigens of interest. This
method was used to clone a diverse panel of human mAbs in
Fab format to tetanus toxoid after generating an antibody library
from peripheral blood lymphocytes isolated from human donors
who had received a booster vaccination.73,74 A major drawback of
this method is its limitation to screening as opposed to selecting
antibody libraries. The number of independent clones, i.e., lysis
plaques, that can be screened, even if the antigen of interest is in
unlimited supply, is practically confined to 106.
Subsequent combinatorial strategies, e.g., phage, yeast and
mammalian cell display, replaced screening with selection, allow-
ing the mining of much larger antibody libraries approaching 1011
independent clones. Selection is facilitated by display technologies
that physically link a displayed antibody fragment to its cDNA
in a defined particle such as a filamentous phage, a ribosome, or
a cell. Millions to billions to, theoretically, trillions of indepen-
dent particles constitute an antibody library that can be mined
with antigens of interest. Particles that display antibody frag-
ments with high affinity to the antigen are isolated. Because of
the simultaneous isolation of their cDNA, the displayed antibody
fragments can be copied, enabling multiple rounds of selection.
In other words, the physical linkage of phenotype and genotype
effectively links recognition and replication. In addition, DNA
sequencing may readily identify phenotype and genotype.
Phage display. The generation and selection of peptide librar-
ies displayed on filamentous phage marked the advent of display
technologies.75 The use of phage display for the generation and
selection of antibody libraries, displayed in either scFv76-78 or
Fab79,80 format, revolutionized the field of mAbs and antibody
engineering in the early 1990s, arguably accelerated by an
intense competition between researchers at the Medical Research
Council (Cambridge, UK) and The Scripps Research Institute
(La Jolla, California, USA). Over the past 20 years, phage display
has proven to be the most robust and versatile mining tool for
human antibody repertoires, yielding human mAbs to virtually
any antigen of interest.8 The process of multiple rounds of selec-
tion on a purified antigen or on antigen-expressing cells, referred
to as panning, can be adapted to positively or negatively select
a range of desired antibody properties, such as affinity, specific-
ity, manufacturability and catalytic activity. While scFv (~25
kDa) and Fab (~50 kDa) still dominate the phage display format,
human antibody repertoires have also been mined through the
display and selection of single variable domains (~12.5 kDa) of
heavy and light chain.81,82
Typically, recombinant fusion to the N-terminus of phage
protein pIII or a fragment of pIII provides the physical linkage
of antibody fragment and phage, facilitating monovalent display
in phagemid systems and multivalent display in phage systems.83
Other multivalent display systems use phage proteins pVIII and
pIX as fusion partners.79,84 For the generation of human mAbs
with high affinity, monovalent Fab display through phage protein
pIII is generally preferred.85 Phage display relies on the expression
Volume 2 Issue 4
and folding of antibody fragments in the periplasm of gram-
negative bacteria, typically Escherichia coli. Although this com-
partment provides an oxidizing milieu suitable for disulfide bridge
formation and mimics the endoplasmic reticulum of eukaryotic
cells surprisingly well, prokaryotic factors that skew the select-
able diversity of antibody libraries are inevitable. For example,
the lack of chaperones in bacteria can lead to inadequate folding
of scFv and Fab and result in toxicity.
Human antibody libraries displayed on phage were generated
from immune and naïve repertoires at about the same time. The
first human immune repertoires mined by phage display were
based on peripheral B cells and bone marrow of vaccinated or
infected human donors and yielded human mAbs to tetanus tox-
oid, HIV-1 gp120, and other viral antigens.85-89 Notably, antibody
libraries derived from bone marrow of HIV-1-positive human
donors with serum antibodies against a variety of other viruses
yielded human mAbs to each of these viruses following selection
by phage display.90 The bone marrow was conceptualized as the
central bank for antigen-experienced B cells that carried a record
of antibody responses to past viral exposures. Autoimmune rep-
ertoires from human donors who have serum antibodies to DNA
and other autoantigens have also been accessed by phage dis-
play.91-93 Tumor-infiltrating B cells provide a unique immune rep-
ertoire in cancer patients that has been mined with phage display
for potentially therapeutic human mAbs to tumor antigens.94-96
Alternatively, peripheral B cells from cancer patients vaccinated
with autologous tumor cells have been used to generate and select
antibody libraries by phage display.97 Peripheral B cells and phage
display were also employed to mine an alloimmune repertoire of
a cancer patient who had been treated with allogeneic hematopoi-
etic stem cell transplantation, revealing serum antibodies to
tumor antigens that might have potential as therapeutic human
mAbs.98
The first human naïve repertoire mined by phage display was
based on peripheral B cells and yielded human mAbs to a vari-
ety of different antigens that had not been encountered in vivo.99
Importantly, this method allowed the generation of human
mAbs to human antigens.100 These studies demonstrated that the
random combination of heavy and light chain can yield human
mAbs with new specificities. Due to the overlap of naïve and
immune repertoires, as well as the bias of mRNA from antibody
secreting cells, antibody libraries from naïve repertoires typically
contain a mixture of heavy and light chain cDNAs with and
without somatic hypermutations. Human mAbs derived from
naïve repertoires generally have lower affinities compared to
human mAbs from immune repertoires, but this shortcoming
can be overcome by increasing the number of independent clones,
thereby generating a larger antibody library.101-104 However, it is
now common to mature the affinity of human mAbs derived
from naïve repertoires with subsequent antibody engineering
methods that are also based on display technologies. Methods
that involve the randomization of certain sequences within heavy
or light chain are discussed in a subsequent subsection on syn-
thetic repertoires.
A method that preserves natural V HDJH, VkJk and VlJl
sequences and yields human mAbs indistinguishable from
www.landesbioscience.com mAbs
endogenous human antibodies involves the shuffling of heavy
and light chains.105 This method, known as chain shuffling, can
yield excellent human mAbs from moderately diverse antibody
libraries with 109 independent clones.106 Chain shuffling can also
be used to convert mouse mAbs to human mAbs via sequen-
tial replacement of mouse heavy and light chains with human
heavy and light chain libraries, respectively, followed by selection
through phage display.107 Adalimumab, the first human mAb
approved by FDA and EMA (Table 1), was derived from a mouse
mAb to human TNFa via this method.108 A similar method,
also based on phage display, preserves only the complementarity
determining region (CDR) 3 sequences of mouse heavy and light
chains, i.e., HCDR3 and LCDR3, while converting everything
else to human sequences.109
A general concern about the de novo generation of human
mAbs from naïve and synthetic repertoires in vitro is their lack of
exposure to negative selection processes in vivo which are integral
to immune tolerance mechanisms and efficiently remove undesir-
able off-target reactivities. Addressing this concern, phage display
has also been used to mine non-combinatorial antibody librar-
ies, i.e., antibody libraries that retain original heavy and light
chain pairings of human antibody repertoires through in-cell
RT-PCR.110 This method is of particular interest for the identifi-
cation of human antibodies in autoimmune repertoires.
Yeast display. The expression, folding and display of antibody
fragments on the surface of cells from the yeast Saccharomyces
cerevisiae involve eukaryotic processes that are similar to those
encountered in the natural B-cell environment. Antibody frag-
ments are channeled through endoplasmic reticulum and Golgi
apparatus, ensuring proper disulfide bridge formation and
N-linked glycosylation.111,112 Thus, yeast display may address
some of the above mentioned biases prokaryotic systems impose
on antibody libraries. In fact, in a direct comparison based on the
same human antibody repertoire, yeast display yielded a more
diverse set of human mAbs than phage display.113 The physical
linkage of antibody fragment and cell is provided through recom-
binant fusion to the C-terminus of the secreted protein Aga2,
which covalently associates with the cell surface protein Aga1.114
Formats of antibody fragments that have been used for yeast
display include scFv,111 Fab115 and scFab.116
Initially limited by the number of independent clones that
could be transformed, yeast display has become a powerful
mining tool for large antibody libraries from human naïve rep-
ertoires.117-119 In addition, yeast display has been successfully
employed for the mining of human immune repertoires.113,120
Interestingly, the haploid/diploid lifecycle of yeast can be
exploited in recombinatorial strategies that generate large anti-
body libraries in diploid cells by mating a haploid cell pool har-
boring a heavy chain library with a haploid cell pool harboring a
light chain library.118,120 Such recombinatorial strategies based on
yeast display have also been used for chain shuffling.121
In contrast to phage display, antibody libraries displayed on
yeast cells are simultaneously screened and selected with an anti-
gen of interest. Screening is carried out by flow cytometry and
selection by FACS. The ability to detect and sort individual bind-
ing events during the selection process has made yeast display
371
an exceptional method for the affinity maturation of mAbs,122
including human mAbs derived from naïve repertoires by phage
display.115
Mammalian cell display. A screening/selection system based
entirely on expression of human antibodies in their natural envi-
ronment, i.e., the mammalian cell, may be best suited for the
mining of human antibody repertoires. Within this system, all
necessary components for human antibody synthesis and process-
ing are available at physiological levels. Compared to yeast cells,
antibody repertoires expressed in mammalian cells are less biased
by properties other than antigen binding. Several laboratories
have therefore developed strategies for human mAb mining by
mammalian cell display. The most straightforward approach is
to transfect an antibody library with a vector directing antibody
expression to the cell surface by means of a C-terminal trans-
membrane region, thereby allowing for selection of cells express-
ing specific antibody based on antigen binding, e.g., by FACS.
In a proof-of-concept study involving the expression of a mix-
ture of two different CD22-specific scFv on the surface of human
embryonic kidney 293T cells, it was estimated that an up to
240-fold enrichment can be achieved with only 2-fold affinity
differences.123 It remains to be shown whether this approach will
allow for isolation of antibodies from more complex mixtures,
such as a library. A general limitation of screening by plasmid
transfection is the potential to deliver multiple plasmids per cell,
leading to the expression of an ill-defined number of different
antibodies on each cell. It would then require multiple rounds
of selection in order to enrich for specific antibodies, even if the
library diversity is small. This problem can be circumvented with
viral expression vectors. As infection rates can be dosed precisely,
conditions can be used where one cell expresses exactly one anti-
body species.
One viral system suitable for screening of human antibody
libraries on the surface of mammalian cells is based on vaccinia
virus.124 Using separate heavy and light chain libraries, African
green monkey kidney BSC-1 cells are co-infected, human IgG
antibodies expressed on the cell surface and cells encoding spe-
cific antibody isolated by FACS. Because heavy and light chains
are expressed from separate vectors, infection rates are kept high
to ascertain that a substantial fraction of cells expresses func-
tional antibody. As a consequence, some of the cells isolated will
express more than one heavy and light chain pair, and several
rounds of selection are beneficial. After screening, specific recom-
binants can be recovered from small numbers of selected cells,
perhaps even single cells.124
A two-step method for the isolation of human antibodies by
mammalian cell display has been described recently.125 Using
virus like particles with highly repetitive arrays of antigen,126
pools of B cells specific for an antigen of interest are first isolated
from peripheral blood lymphocytes of immunized or naturally
immune human donors by FACS. Recombinant, antigen-focused
scFv libraries are then generated from these B cells and screened
by mammalian cell surface display using a Sindbis virus expres-
sion system. Infection of baby hamster kidney cells at a low
multiplicity of infection ensures that one cell expresses only one
antibody species, thus allowing identification of antigen-specific
372 mAbs
antibodies in a single round of FACS. Due to the highly repli-
cation-competent nature of the Sindbis vector, viruses encoding
a specific antibody can readily be amplified from single-sorted
cells. Thus, the binding properties of antibodies can be charac-
terized prior to cloning, sequencing and subsequent production
as whole IgG. Towards a clinical application of the technology,
therapeutic mAbs against nicotine and influenza A M2 protein
have been isolated, characterized and validated preclinically to
date.125,127
A drawback of mammalian cell display is that the number
of cells that can be handled at the same time is limited. This is
particularly true if the screening is done under conditions where
one (or close to one) antibody species is expressed per cell, limit-
ing the diversity of libraries that can be conveniently handled to
about 107 independent clones. This is suboptimal for combinato-
rial libraries and pre-selection of B cells for antigen specificity is
advisable. It is therefore no coincidence that the human mAbs
isolated by mammalian cell display to date are derived from pools
of B cells enriched for antigen specificity.125,127 Because these
enriched combinatorial libraries are of low diversity, there is a
high likelihood that the resulting human mAbs contain natural
pairs of heavy and light chains, i.e., are derived from clonally
related B cells. An advantage common to all screening systems
based entirely on mammalian cell expression is that they have
a built-in selection for efficient expression in mammalian cells.
As antibody good manufacturing practice (GMP) processes typi-
cally involve expression in mammalian cells, this is expected to
have a significant impact on the cost of goods.
Non-Natural Repertoires
Transgenic repertoires. More than 20 years ago, a proof of con-
cept study first demonstrated that human Ig gene segments intro-
duced into a mouse can be accessed by the mouse immune system
and used to express functional antibodies.128 In this approach, a
human heavy chain mini locus containing V H, D and JH gene
segments linked to a C m gene was introduced into an embry-
onic stem cell that was used to develop a transgenic mouse. In
this mouse, a large fraction of lymphoid cells expressed human
antibody genes and this led to measurable levels of hybrid IgM
with human heavy chains and mouse light chains in the serum.
Further, it was shown that conventional hybridoma technology
could be used to recover recombinant monoclonal IgM, thus lay-
ing the ground for the production of human antibodies in so
called humanized mice.
Production of human antibodies in genetically modified mice
was immediately recognized as a highly attractive approach for
the generation of human mAbs against human antigens, as mice
can readily be immunized with any antigen of interest and are not
tolerant to most human proteins.129 While the mice described by
Brüggemann et al.128 only expressed a human heavy chain mini
locus in the presence of the endogenous mouse heavy and light
chain loci, the production of fully human antibodies became
feasible with the development of mice with additional genetic
modifications. In two different laboratories, mice were generated
that combined disruptions of the endogenous mouse heavy and
Volume 2 Issue 4
kappa light chain loci and transgenes encoding human heavy and
kappa light chain loci.130,131 The endogenous mouse lambda light
chain locus was left unmodified by both groups because it only
contributes to 5–15% of the antibody repertoire.132 In the report
by Lonberg et al.131 the human heavy chain locus comprised
3 V H, 16 D, all 6 JH and the C m and C g1 constant region gene seg-
ments, whereas the light chain locus comprised 4 Vk, all 5 Jk and
the C k gene segments. The mice described by Green et al.130 car-
ried a repertoire of similar diversity, with the heavy chain locus
including 5 V H, all 25 D, all 6 JH and the C m and C d constant
region gene segments, and the light chain locus including 2 Vk,
all 5 Jk and the C k gene segments. The human transgenes in such
mice had undergone V HDJH and VkJk rearrangements with N
region diversification, i.e., additions or deletions of nucleotides at
the V H-D-JH and Vk -Jk junctions, class switch recombination and
somatic hypermutation.131 Despite the limited repertoire present
in both transgenic mouse strains, immunization led to the gen-
eration of specific human antibodies against several antigens that
were accessible by conventional hybridoma technology.130,131 In
this respect, transgenic mice have led to a revival of hybridoma
technology for the mining of human antibody repertoires.
The finding that transgenic mice containing human heavy
and light chain mini loci are capable of mounting a specific
humoral immune response comprising human antibodies is sur-
prising in at least two respects. First, it demonstrates that hybrid
B-cell receptor complexes consisting of human surface Ig and
mouse Iga/Igb (CD79a/CD79b) chains are functional in vivo
and permit largely normal development and function of B cells.
Second, only a fraction of the natural heavy and light chain V
gene repertoire appears to be sufficient for the generation of
antigen-specific antibodies, putting the relative importance of
combinatorial diversity into perspective. For the production of
antibodies against protein antigens using hyperimmunization
protocols, the two non-germline encoded sources of clonal diver-
sity, i.e., junctional diversity and somatic hypermutation, appear
to be sufficient. Indeed, it was shown that the HCDR3 repertoire
is in large part created by junctional diversity, and sufficient for
most antigen specificities.133
Despite the initial success of the transgenic mice carrying
mini loci, human mAbs against few antigens were isolated from
these mice. The immune responses in these mice were inferior
to those of wild type mice, with reduced numbers of mature B
cells and lower levels of circulating Igs; however, incorporation
of a significantly larger repertoire of Vk gene segments led to
increased levels of pre-B and B cells.134 Mice that incorporated
the majority of the human heavy chain and kappa light chain loci
were found to recapitulate the human antibody response even
better by restoring B-cell compartments to near normal levels.135
Compared to the transgenic mice carrying mini loci,130,131 the
animals with near complete loci displayed higher levels of human
serum Ig, better class switch recombination efficiency and
increased clonal diversity and magnitude of the human antibody
response. Accordingly, a panel of human mAbs with high affinity
against three human antigens was isolated.135 In contrast to mice,
approximately 40% of serum Igs in humans contains a lambda
light chain. To further enhance the diversity of humanized B-cell
www.landesbioscience.com mAbs
responses, transgenic mice were generated that carried, for the
first time, large portions of all three human Ig loci, including the
human lambda light chain locus, in a background in which the
endogenous mouse heavy and kappa light chain loci had been
inactivated. Upon immunization, these mice generated antibod-
ies containing both human kappa and lambda light chains.136
To generate more complete humanized mice, the to date larg-
est fraction of the human germline repertoire was introduced by
microcell-mediated chromosome transfer. In this manner, tran-
schromosomal mice were generated that express the complete
heavy chain and kappa light chain repertoire in absence of mouse
heavy and kappa light chains.137 However, instability of the chro-
mosome fragment containing the kappa light chain locus led to a
substantial reduction in the generation of hybridomas, a problem
that was solved by crossing IgH transchromosomal mice with Igk
transgenic mice.138 It is interesting to note that of the six human
mAbs on the market today, five are derived from humanized
mice (Table 1). Of these, panitumumab is derived from Abgenix’
XenoMouse,135 whereas the other four are derived from Medarex’
UltiMab platform, which is based on the HuMab mouse,134 the
Kirin TC mouse137 and a cross of the two, the KM mouse.138
Natural antibody responses are polyclonal. There are cases
in which immunotherapy using polyclonal antibodies (pAbs)
instead of mAbs is expected to be more efficient. This includes,
for example, the treatment or prevention of diseases caused by
different viral or bacterial strains or combating viruses prone to
formation of escape mutants. In principle, human pAbs could
be produced in humanized mice.138 While mice are well suited
for the generation of mAbs, their small body size makes them
unsuitable for the production of useful amounts of therapeutic
or preventive pAbs. One method of accessing the potential of
human pAbs is to use recombinant pAbs, i.e., defined cocktails
of mAbs.139 In addition, due to advances into the understand-
ing of humoral immune systems of large farm animals combined
with significant technological advances, it may soon be possible
to generate human pAbs by hyperimmunization of transchromo-
somal cattle.140
Synthetic repertoires. Antibody libraries from synthetic rep-
ertoires are based on synthetic DNA sequences designed to diver-
sify the antibody sequence.69,141 Depending on the design, human
mAbs from synthetic repertoires may or may not be distinguish-
able from endogenous human mAbs. The first human synthetic
repertoire was generated by randomizing all of the 16 amino
acids that comprised the HCDR3 sequence of a human anti-
tetanus toxoid mAb.142 Subsequent selection by phage display
against fluorescein yielded a panel of human mAbs that revealed
a shift in specificity from tetanus toxoid to fluorescein. In addi-
tion, synthetic repertoires allow the CDR grafting of a peptide
motif with known specificity followed by randomization of the
flanking sequences and selection by phage display. This method
yielded human mAbs that bind human integrins aVb3 and aIIbb3
with high affinity.143,144 The randomization of single or multiple
CDR sequences followed by selection with phage display has also
been utilized for the affinity maturation of human mAbs that
were derived from immune or naïve repertoires. This process can
yield human mAbs with picomolar affinity.145,146
373
 Antibody libraries that are based on human naïve repertoires
but introduce additional sequence diversification through ran-
domizing single or multiple CDR sequences represent, by defini-
tion, human synthetic repertoires;147-150 however, not all synthetic
repertoires are based on randomized CDR sequences. Using mas-
ter framework sequences derived from a single human V H and Vl
gene segment, Söderlind et al.151 introduced and recombined a
complete set of CDR sequences derived by RT-PCR from human
naïve repertoires. Yet another approach of sequence diversification
in vitro is based on dispersing mutations throughout heavy and
light chain variable domain sequences, as opposed to focusing
mutations on defined segments such as CDR sequences. This can
be achieved through error-prone PCR or mutagenic Escherichia
coli strains. Like focused mutagenesis, dispersed mutagenesis has
been utilized for the affinity (or catalytic activity) maturation of
human mAbs, usually by phage display152,153 and, more recently,
by yeast display.115,154
Owing to the fact that the discussed antibody libraries blend
in vivo and in vitro diversity, they are sometimes referred to as
semi-synthetic. By contrast, fully synthetic antibody libraries
are entirely designed and engineered in vitro. A fully synthetic
antibody library that combined a single human V H and Vk gene
segment with randomized HCDR3 and LCDR3 sequences
was mined successfully for human mAbs to human antigens.155
Further restrictions that mimic natural CDR sequence diversity
in the heavy chain variable domain and use a fixed light chain
variable domain yielded human mAbs that closely resemble
endogenous human antibodies. These were improved further by
introducing natural CDR sequence diversity in the light chain
variable domain.156,157 CDR sequence diversity can be further
restricted to a four-amino-acid code (Ala, Asp, Ser and Tyr)158
or even a two-amino-acid code (Ser and Tyr),159 demonstrating
the versatility of synthetic repertoires and providing molecular
insights into specificity and affinity of antibody/antigen interac-
tions. Restricting sequence diversity in fully synthetic and semi-
synthetic repertoires has also been used for tailoring antibody
libraries toward the selection of human mAbs to a particular
class of antigens, such as peptides and carbohydrates.160,161 Fully
synthetic antibody libraries that are more complex and based on
>1010 independent clones (in scFv or Fab format) reassemble a
variety of natural CDR and framework sequences and combine
these with randomized CDR sequences.162,163 A fully synthetic
antibody library with diversification in all six CDR sequences
was reported more recently.164
In addition to phage display, fully synthetic antibody libraries
have been mined successfully with ribosome display.165 Ribosome
display is an in vitro display technology166 that does not require
transformation of prokaryotic or eukaryotic cells.167-169 Ribosome
display has been employed to both accomplish and guide the
affinity maturation of human mAbs.170 Other less common dis-
play technologies, such as retroviral display,171 have also been used
for mining synthetic repertoires. In addition to the conventional
scFv and Fab formats, synthetic repertoires have facilitated the
display and selection of single human variable domains 81,172 and,
recently, human constant domains with diversified sequences in
non-CDR loops of the Ig fold.173,174
374 mAbs
Another exciting advance in the area of human synthetic rep-
ertoires has been the generation and selection of antibody librar-
ies that feature an expanded genetic code for the incorporation
of unnatural amino acids into CDR sequences.175 Collectively,
synthetic repertoires have begun to outshine all other human
antibody repertoires with respect to diversity and versatility.
Human mAbs from synthetic repertoires are poised to become
preferred reagents for diagnostic and basic applications; however,
their utility for preventive and therapeutic interventions remains
to be established in clinical trials.
Outlook
Increased availability and improved accessibility of human anti-
body repertoires have made human mAbs the entity of choice for
therapeutic mAbs. Today, human mAbs to virtually any antigen
of interest can be generated. The clinical performance and the
commercial viability of currently approved human mAbs provide
a robust platform for the development of future generations of
human mAbs. Not only will these broaden the scope of targeted
antigens and indications, but also improve the activity of inter-
vention through better pharmacodynamic and pharmacokinetic
properties. Although an initial goal in the development of human
mAbs was to render them indistinguishable from endogenous
human antibodies, subtle antibody engineering and expression
strategies that retain low immunogenicity, but tune affinity,
effector functions and circulatory half-life, as well as lower the
cost of goods, are being employed in the development of the next
generation of human mAbs.176
Of the currently approved human mAbs (Table 1), one was
derived from a naïve repertoire and five were derived from trans-
genic repertoires. It will be interesting to compare human mAbs
from different sources with respect to their pharmacokinetic and
pharmacodynamic performance. For example, the human anti-
human TNFa mAb, adalimumab, which was derived indirectly
from a naïve repertoire as described above, induces a HAHA
response in a significant percentage of patients, adversely affect-
ing serum concentrations and clinical responses.177 The recent
approval of human anti-human TNFa mAb, golimumab,178
which was derived from a transgenic repertoire, provides a first
opportunity to clinically investigate whether human mAbs
derived from different human antibody repertoires against the
same antigen differ in terms of immunogenicity.
The immune repertoire is a natural repertoire that has not been
fully exploited for the mining of human mAbs. Sophisticated in
vivo processes that shape the immune repertoire afford endog-
enous human antibodies with high affinity, minimal immunoge-
nicity and minimal off-target reactivity. The immune repertoire
has become increasingly accessible through new methods that
facilitate the enrichment, isolation or expansion of antigen-spe-
cific post-GC B cells, plasma blasts, plasma cells or memory B
cells from peripheral blood. The development of these methods
has been spurred by the discovery of endogenous human antibod-
ies of potential therapeutic utility in normal human donors, as
well as in those with inflammatory diseases, infectious diseases,
and cancer. Endogenous human antibodies that are triggered
Volume 2 Issue 4
by preventive or therapeutic interventions, such as vaccination
or allogeneic hematopoietic stem cell transplantation, are par-
ticularly attractive. An ideal mining strategy for this exceptional
source of human mAbs is mammalian cell display as it provides
an environment that closely resembles B cells. Consequently, this
platform is anticipated to contribute to the development of future
generations of human mAbs.
Finally, it is important to keep in mind that endogenous
human antibodies are polyclonal whereas human mAbs, by
definition, are monoclonal. In general, polyclonal or even oligo-
clonal responses are thought to be more proficient in mounting
effector functions compared to monoclonal responses.179 While
non-recombinant human pAbs purified from human donors
have long been used in preventive and therapeutic interventions,
recombinant human pAbs would afford much broader applica-
bility.139 Thus, despite current practical and regulatory challenges
in manufacturing and formulation, human pAbs will likely be a
Conflict-of-interest and financial disclosure
statements
R.R.B. is an employee of Cytos
Biotechnology AG and holds stock options
in the company.
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