ANTIBODIE

S
STRUCTURE CLASS
POLYCLONAL ANTIBODIES
PRODUCTION
ANTIBODIE
S
Derived from
diferent B
Lymphocytes cell
lines
OL!CLONAL" #ONOCLONAL"
Derived from $ sin%le
B cell clone
B$tch to B$tch
v$ri$tion $fectin%
A& re$ctivity ' titre
mA& ofer
Reprod(ci&le)
redict$&le '
otenti$lly
ine*h$(sti&le s(pply
of A& +ith e*,(isite
speci-city
En$&le the
development of
sec(re imm(no$ss$y
systems"
NOT o+erf(l tools
for clinic$l di$%nostic
tests
Aminopterin blocks DNA
synthesis by e no!o
p"th#"y
DENOVO PATHWAY
Phosphoribosyl pyrophosphate
+
Uridylate
Aminopterin
Nucleotides
DNA
A!VA"E PATHWAY
Hypo#anthin
e
H"P$T+
Hypo#anthin
e %uanine
phosphoribos
yl
trans&erase
Thymidine
T'+
(thymidine )inase*
$ybriom" technolo%y
Nobel Pri&e

"eor%es '+hler, -.sar /ilstein, and

Niels 'a0 1erne in 2345 shared the
Nobel Pri6e in Physiolo%y or /edicine
in 2378 &or the disco9ery:
ELISA screenin% 'or
monoclon"l "ntiboy 'or
speci'ic "ntiboy
The ;ells are coated ;ith the appropriate $nti%en:
The solution (e:%:, serum* containin% antibodies is added:
A&ter they ha9e had time to bind to the immobili6ed
$nti%en,

an en6yme<con0u%ated $nti.imm(no%lo&(lin is added,

consistin% o&

an antibody a%ainst the antibodies bein% tested &or: =or

e#ample, i& human anti<H>V antibodies are bein% assayed,
then antibodies (raised in a %oat or rabbit a%ainst human
immuno%lobulins* are con0u%ated to the en6yme:
A&ter ;ashin% a;ay unreacted rea%ent, the substrate is
added:

The intensity o& the color produced is proportional to the

amount o& en6yme<labeled antibodies bound (and thus to
the concentration o& the antibodies bein% assayed*:
$(m"n monoclon"l
"ntiboies

Problems

!ac) o& human myeloma cells (immortal

%ro;th, H"P$T<, not secrete ab, and support
ab production in them*

Alternati9e ? cell immortali6ed by

trans&ormation ;ith E?V*

Another problem di@culty to %et anti%en

acti9ated ? cells &rom human lymphoid
tissues

One ;ay cultured human cells ;ith anti%en

(produce lo; a@nity >%/ Ab

Or Human cells in ->D mice:

Applic"tions o'
)onoclon"l Antiboies

Dia%nostic Applications

Therapeutic Applications

-linical Applications
Imm(noto*ins
C"t"lytic monoclon"l
"ntiboies +Ab&ymes,

An ab6yme (&rom antibody and en6yme*, also called catmab

(&rom catalytic monoclonal antibody*, is a monoclonal antibody
;ith catalytic acti9ity:

En6ymes &unction by lo;erin% the acti9ation ener%y o& the

transition state, thereby cataly6in% the &ormation o& an other;ise
less<&a9orable molecular intermediate bet;een reactants and
products: >& an antibody is de9eloped to a stable molecule thatAs
similar to an unstable intermediate o& another (potentially
unrelated* reaction, the de9eloped antibody ;ill en6ymatically
bind to and stabili6e the intermediate state, thus cataly6in% the
reaction: A ne; and uniBue type o& en6yme is produced:

central %oal o& catalytic antibody research is the deri9ation o& a

battery o& ab6ymes that cut peptide bonds at speciCc amino acid
residues, much as restriction en6ymes cut DNA at speciCc sites:
-DA "ppro!e ) Ab
ther"py
Why should we be
interested ?

mAbs drive the development of multibillion dollar

biotechnology industry.

Many of the leading pharmaceutical companies

have entered the mAb sector, attracted by
quicker and less costly development, higher
success rates, premium pricing.

The outlook for monoclonal antibody therapeutics

is healthy. The ongoing success of existing
products, combined with a bulging pipeline of
new products awaiting approval and limited
generic erosion, point towards robust growth in
this segment