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Mechanism 1: Tagging
Mechanism 2: Neutralization
Mechanism of
Antibodies (Tagging)
3) Stimulates
2) Antibody
other immune
binds to the 4)Antigen is
1) Antigen is responses
antigen removed
detected such as
(Antigen is from the body
complement
tagged)
pathway
Mechanism of
Antibodies
http://www.youtube.com
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Development of
Monoclonal Antibodies
ANTIGENS
IMMUNISATION
HOST
The first step in making a
ANIMAL hybridoma is to generate antibody-
(MOUSE) producing B lymphocytes
(immune cells of a mouse).
Multiple immunizations are
performed until an appropriate
level of antibodies are achieved.
TEST
BLEEDS
Myeloma cells
Spleen of mouse
Polyethylene glycol
(PEG) is added to culture
to promote membrane
fusion.
Hybridomas
Aminopterin
Thymidine
Unfused myeloma
cells
THEREFORE, only the
Unfused spleen cells hybridomas survive as
the myeloma cells cannot
Hybridomas
replicate their DNA (
because they lack
HGPRT) and unfused
spleen cells have limited
lifespan.
Surviving
Hence the
hybridomas
name,
are separated
monoclonal,
and
as the cells
individually
are derived
cultured. (one
from one
cell per wall)
single cell.
After a few weeks of culture,
the culture fluid is screened
for presence of desirable
antibodies.
1) Phage display-generated
antibodies
Use of bacteriophage and E.
coli
2) Transgenic Mice
Insert human antibody
gene loci
knocking out genes for
making antibodies
Uses of monoclonal
antibodies
Diagnostic tests
Used in several diagnostic tests to detect small amounts
of drugs, toxins, hormones or infectious diseases.
1) Immunoassay procedures
2) Radioimmunodetection - attachment of MAbs to
radioactive atoms
3) Fluorescent molecules or metal atoms such as copper
and gold coupled to the antibody to assist in imaging
the target.
Diagnostic tests
Monoclonal antibodies to human chorionic
gonadotropin (HCG) are used in pregnancy test kits.
Diagnosis of AIDS by the ELISA (enzyme-linked
immunosorbent assay) test
Monoclonal antibody
therapy
Use of monoclonal antibodies to specifically bind to
antigens of target cells or proteins. This may then
stimulate the patient's immune system to attack
those cells.
Monoclonal antibodies may soon be extended to the
therapeutic treatment of diseases such as infectious
diseases and ophthamology.
With sales exceeding 32 billion dollars in 2008, MAbs
are set to become an important sector of the
pharmaceutical industry.