MONOCLONAL

antibodies

Dr.T.V.Rao MD
Beginning of Monoclonal Era
 Georg Kohler and Cesar Milstein
fuse mouse lymphocytes with
neoplastic mouse plasma cells to
yield hybridomas that produce
specific antibodies. This offers a
limitless supply of monoclonal
antibodies. Monoclonal antibodies
permit diagnostic tests that are
specific, and function as probes.
 Discovery of Monoclonal
Antibodies
 Monoclonal
antibodies were
produced in mice
using a technique
described by
Köhler and
Milstein et al..
They were awarded
the Nobel Prize in
1984 (along with
Jerne) for their
work.
Nobel prize in Medicine and Physiology
was awarded to Köhler, Milstein and
Jerne in 1984
 Monoclonal Antibodies
 Monoclonal antibodies are
Monospecific antibodies that are
identical because they are produced
by one type of immune cell that are
all clones of a single parent cell.
Given (almost) any substance, it is
possible to create monoclonal
antibodies that specifically bind to
that substance
 Nomenclature
 The nomenclature of monoclonal
antibodies is a naming scheme for
assigning generic, or non-proprietary,
names to a group of medicines called
monoclonal antibodies. This scheme
is used for both the World Health
Organization’s International Non-
proprietary Names and the United
States Adopted Names
 Study of Myeloma leads to
Discovery of Monoclonal
antibodies
 In the 1970’s the B-
cell cancer myeloma
was known, and it was
understood that these
cancerous B-cells all
produce a single type
of antibody. This was
used to study the
structure of
antibodies, but it was
not possible to
produce identical
antibodies specific to
a given antigen.
Fusion of Mice spleen cells with
Myeloma cells produced
Monoclonal antibodies
 Characters of Monoclonal
Antibodies
 Monoclonal antibodies (mAb) are
a single type of antibody that are
identical and are directed against a
specific epitope (antigen, antigenic
determinant) and are produced by B-
cell clones of a single parent or a
single hybridoma cell line. A
hybridoma cell line is formed by the
fusion of a one B-cell lymphocyte
with a myeloma cell. Some myeloma
cells synthesize single mAb
antibodies naturally
Hybridoma creates Monoclonal
antibodies
 Monoclonal antibodies
are typically made by
fusing myeloma cells
with the spleen cells
from a mouse that has
been immunized with
the desired antigen.
However, recent
advances have
allowed the use of
rabbit B-cells.

Doctorrao’s ‘e’ learning

 Producing Monoclonal
antibodies
 First, a mouse is
inoculated with the
antigen to which
the MA are to be
produced. After
this, the spleen is
removed and fused
with myeloma cells
in order to produce
the hybridomas
that will be selected
according to the
antibody produced.
Hybridoma leads to
Proliferation
 Principles in Hybridoma
technology
 inject the protein into a
mouse.
 - remove the spleen.
 - identify which spleen
cells are producing
antibodies.
 - separate these cells
and grow in tissue
culture tubes.
 - screen each Ab for
cross reactivity.
 - select the Ab which
doesn't cross react with
any other protein.

Doctorrao’s ‘e’ learning series

 Monoclonal antibodies are
produced by Hybridoma technique
Monoclonal – Diagnostic use
 Although monoclonal
antibodies were first
produced in 1975 as
research tools,
scientists quickly
recognized their
practical uses,
especially in
diagnostic tests and in
therapy. Several
diagnostic procedures
that use monoclonal
antibodies are now
available
Doctorrao’s ‘e’ learning series
A breakthrough in
Diagnostics
 A monoclonal
antibody can be used
to detect
pregnancy only
14 days after
conception. Other
monoclonal antibodies
allow rapid diagnosis
of hepatitis,
influenza, herpes,
streptococcal, and
Chlamydia
infections.

Doctorrao’s ‘e’ learning

 Helps in Critical Diagnostic
decisions
 They can be used
to detect for the
presence and
quantity of this
substance, for
instance in a
Western blot test
(to detect a
substance in a
solution) or an
immunofluorescenc
e test.
Doctorrao’s ‘e’ learning series
 Monoclonal's helps In
Immunodiagnostic tests
 Monoclonal
antibodies can
also be used to
purify a
substance with
techniques
called
immunoprecipi
tation and
affinity
Limitations with Mouse
Monoclonals
 Problem in medical
applications is that
the standard
procedure of
producing
monoclonal
antibodies yields
mouse antibodies,
and these are
rejected by the
human immune
system
Finding solutions for Human
use
 In one approach, one
takes the DNA that
encodes the binding
portion of monoclonal
mouse antibodies and
merges it with human
antibody producing
DNA, in order to make
bacteria produce
antibodies that are
half mouse and half
human.

Doctorrao’s ‘e’ learning

series
 Conjugated monoclonal
antibody therapy:
 Toxins or radioactive
isotopes are bound to
the constant region of
the MAbs. When the
MAb binds to the
surface cells of a
tumor the toxin or
radioactivity will kill
the cancer cells and
all cells within a
certain radius (a killing
zone). In this way
cancer cells within the
tumor will be killed

Doctorrao’s ‘e’ learning series

 Knowledge on Monoclonal’s
advances
 Mice have been genetically engineered to
produce antibodies that have human
constant regions (this is the part of the
antibody that the human immune system
recognizes as being foreign (mouse)). By
using these hybrid (or chimeric
monoclonal antibodies with human
constant regions, the immune system only
"sees" a human protein and does not react
against them. So, they can be injected
many times to kill all of the cells in a
tumor.
 Monoclonal antibodies for
cancer treatment

 Possible treatment for

cancer involves
monoclonal antibodies
that bind only to
cancer cells specific
antigen and induce
immunological
response on the target
cancer cell (naked
antibodies). mAb can
be modificated for
delivery of [toxin],
radioisotope, cytokine.

Doctorrao’s ‘e’ learning series

 FDA approves
 The example
described in class is
Herceptin. These
monoclonal
antibodies can be
used against certain
forms of breast
cancer and have
passed clinical trials
and been approved
for use by the FDA.
 FDA approves and Trails on
 The first FDA-approved
therapeutic monoclonal
antibody was a murine
IgG2a CD3 specific
transplant rejection drug,
OKT3 (also called
muromonab), in 1986.
This drug found use in
solid organ transplant
recipients who became
steroid resistant.
Hundreds of therapies
are undergoing
clinical trials. Most are
 Created for
Medical and
Paramedical
Students in the
Dr.T.V.Rao MD
DevelopingEmail
world
doctortvrao@gmail.com