7. ANTIBODIES
POLYCLONAL MONOCLONAL
 Derived from different B  Derived from a single B cell
Lymphocytes cell lines. clone.
 Batch to Batch variation  No Batch to Batch
affecting Ab reactivity & variations. Effectiveness of
treatment. Ab is much more
predictable.
 NOT Powerful tools for  Enable the development of
clinical diagnostic tests. secure immunoassay
systems.

9. 1) Homogeneity:
Monoclonal antibody represents a single antibody molecule that
binds to antigens with the same affinity and promote the same
effectors functions.
2) Specificity:
The product of a single hybridoma reacts with the same epitope on
antigens.
3) Immunizing Antigen:
Need not be pure or characterized and is ultimately not needed to
produce large quantities of antibody.
4) Selection:
It is possible to select for specific epitope specificities and generate
antibodies against a wider range of antigenic determinants.
5) Antibody Production:
Unlimited quantities of a single well-defined monospecific reagent.

10. 1) Affinity:
Average affinity of monoclonal antibodies are generally
lower than polyclonal antibodies.
2) Effector Functions:
Because antibody is monoclonal, it may not produce the
desired biologic response.
3) Specificity:
Monoclonals against conformational epitopes on native
proteins may lose reactivity with antigens.
4) Cross reactions:
Antibodies sometimes display unexpected crossreactions
with unrelated antigens.
5) Time and effort commitment:
VERY LARGE.

13. 1) FILTRATION METHOD:
Cells, cell
debris, lipids, and
clotted material are
first
removed, typically by
filtration with a 0.45
um filter.

14. a) Most of the charged impurities are usually
anions such as nucleic acids and endotoxins.
These are often separated by ion exchange
chromatography.
b) column chromatography can also use which is
much quicker method.

15. 2) ANTIBODY HETEROGENEITY:
 Product heterogeneity is common to monoclonal antibody and
other recombinant biological production and is typically
introduced either upstream during expression or downstream
during manufacturing.
 These variants are typically
aggregates, deamidation products, glycosylation variants, oxidized
amino acid side chains, as well as amino and carboxyl terminal
amino acid additions. These seemingly minute changes in a
monoclonal antibody’s structure can have a profound effect on
preclinical stability and process optimization as well as
therapeutic product potency, bioavailability, and immunogenicity.
The generally accepted method of purification of process streams
for monoclonal antibodies includes capture of the product target
with Protein A, elution, acidification to inactivate potential
Mammalian viruses, followed by cation exchange
chromatography, and finally anion exchange chromatography.

16. Recombinant antibody engineering involves the use
of viruses or yeast to create antibodies, rather than
mice. These techniques rely on rapid cloning of
immunoglobulin gene segments to create libraries of
antibodies with slightly different amino
acid sequences from which antibodies with desired
specificities can be selected. These techniques can
be used to enhance the specificity with which
antibodies recognize antigens, their stability in
various environmental conditions, their therapeutic
efficacy, and their detect ability in diagnostic
applications. Fermentation chambers have been
used to produce these antibodies on a large scale.

17. In one approach, mouse DNA encoding the
binding portion of a monoclonal antibody was
merged with human antibody-producing DNA in
living cells, and the expression of
this chimeric DNA through cell culture yielded
partially mouse, partially human monoclonal
antibody. For this product, the descriptive terms
"chimeric" and "humanised" monoclonal
antibody have been used to reflect the
combination of mouse and human DNA sources
used in the recombinant process.

19. A method to generate fully human monoclonal antibodies
using blood from normal donors without previous exposure
to the antigen. The technology uses to generate an antigen
presentation system, which emulates the natural
presentation of antigens by the immune system.

21.  Radio immunotherapy (RIT)
Antibody-directed enzyme prodrug
therapy (ADEPT)
 Immunoliposomes

22. Monoclonal antibodies used for autoimmune
diseases include infliximab and adalimumab, whic
h are effective in rheumatoid arthritis, Crohn's
disease and ulcerative Colitis.
Basiliximab and daclizumab activated T cells and
thereby help preventing acute rejection of kidney
transplants.
Omalizumab inhibits human immunoglobulin E
(IgE) and is useful in moderate-to-severe
allergic asthma.

23. Antibodies are used in several diagnostic tests to
detect small amounts of drugs, toxins or
hormones.
e.g. monoclonal antibodies to human chorionic
gonadotropin (HCG) are used in pregnancy test
kits.
Another diagnostic uses of antibodies is the
diagnosis of AIDS by the ELISA test.

25. Antibody Brand name Type Indication
Abciximab ReoPro chimeric Cardiovascular disease
Adalimumab Humira human Several auto-immune
disorders
Alemtuzumab Campath humanized Chronic lymphocytic
leukemia
Basiliximab Simulect chimeric Transplant rejection
Belimumab Benlysta human Systemic lupus
erythematosus
Colorectal cancer, Age
Bevacizumab Avastin humanized related macular
degeneration
Brentuximab
Adcetris Chimeric Hodgkin lymphoma
Vedotin

26. Antibody Brand name Type Indication
Canakinumab Ilaris Human
Cetuximab Erbitux chimeric Colorectal cancer, Head
and neck cancer
Certolizumab Cimzia humanized Crohn's disease
pegol
Daclizumab Zenapax humanized Transplant rejection
Denosumab Prolia , Xgeva Human Postmenopausal
osteoporosis
Eculizumab Soliris humanized Paroxysmal nocturnal
hemoglobinuria
Efalizumab Raptiva humanized Psoriasis

27. Antibody Brand name Type Indication
Palivizumab Synagis humanized Respiratory Syncytial
Virus
Panitumumab Vectibix human Colorectal cancer
Ranibizumab Lucentis humanized Macular degeneration
Rituximab Rituxan, chimeric Non-Hodgkin
Mabthera lymphoma
Tocilizumab Actemra Humanised Rheumatoid arthritis
Tositumomab Bexxar murine Non-Hodgkin
lymphoma
Trastuzumab Herceptin humanized Breast cancer