This is about the research paper; Continuous cultures of fused cells secreting antibody of predefined specificity.

2. WHO is SHE???
CAN YOU GUESS HER???

3. HENRIETTA LACKS

4. César Milstein & Georges J. F. Köhler
Presented by S.M Gharib Nawaz
Continuous cultures of
fused cells secreting
antibody of predefined
specificity

6. César Milstein:
(8 October 1927 – 24 March
2002) was an Argentine biochemist
in the field of antibody research.
Georges Jean Franz Köhler:
(April 17, 1946 in Munich –
March 1, 1995) was a German
biologist.
César Milstein and Niels Kaj
Jerne, Köhler won the Nobel Prize
in Physiology or Medicine in 1984

7. ANTIBODIES & ANTIGENS:
• Antibodies are proteins produced by the body's immune system in
response to antigens, which are harmful substances.
• Antigens include bacteria, fungi, parasites, viruses, chemicals, and
other substances the immune system identifies as foreign.

8. ANTIBODIES ISOTOPES:
• Human antibodies are classified into five isotypes:
1. IgG is the most abundant in the blood (plasma), accounting for 70-75% . It
detoxifies harmful substances and is important in the recognition of antigen-
antibody complexes. IgG is transferred to the fetus through the placenta and
protects the infant until its own immune system is functional.
2. IgM usually circulates in the blood, accounting for about 10% of human
immunoglobulins. IgM has a pentameric structure in which five basic Y-shaped
molecules are linked together. B cells produce IgM first in response to
microbial infection/antigen invasion.
3. Although IgM has a lower affinity for antigens than IgG, it has higher avidity
for antigens because of their structure. IgM, by binding to the cell surface
receptor, also activates cell signaling pathways.

9. ANTIBODIES ISOTOPES:
3. IgA is abundant in serum, nasal mucus, saliva, breast milk, and
intestinal fluid, accounting for 10-15%. IgA forms dimers (i.e., two IgA
monomers joined together). IgA in breast milk protects the
gastrointestinal tract of neonates from pathogens.
4. IgE is present in minute amounts, accounting for no more than
0.001% of human immunoglobulins. Its original role is to protect against
parasites. In regions where parasitic infection is rare, IgE is primarily
involved in allergy.
5. IgD accounts for less than 1% of human immunoglobulins. IgD may
be involved in the induction of antibody production in B cells, but its
exact function remains unknown.

11. MONOCLONAL & POLYCLONALANTIBODIES:
• Monoclonal antibody means that the man-made antibody is
synthesized from cloned immune cells, and the identical monoclonal
antibody produced binds to one type of antigen.
• Polyclonal antibodies are synthesized from different immune cells
and the antibodies produced bind to multiple antigens.

12. 1. Immunization of an animal.
2. Cell Fusion.
3. Selection of Hybridomas.
4. Isolation of hybridomas of single specificity.
5. Screening of products.
6. Cloning and Propagation.
7. Characterization and Storage.
Production of Monoclonal Antibodies

13. 1. Immunization of an animal:

14. 2. Cell Fusion:
TK
Hypoxanthine Guanosine Phosporibosyl Transferase
Thymidine Kinase

15. 2. Cell Fusion:

16. 3. Selection of Hybridomas:
HAT MEDIUM
Hypoxanthine
Aminopterin
Thymidine

17. De novo pathway
Aminopterin
Salvage pathway
HGPRT
TK
to produce antibodies

18. Switch to salvage but
limited life span
Unable to switch
HGPRT-
Switch to salvage pathway
Unlimited life span

19. 4. Isolation of hybridomas of single specificity:

20. 5. Screening of products:
Two techniques used for this purposes
1. ELISA (Enzyme Linked Immunosorbent Assay)
2. RIA (Radio Immuno Assay)

21. 6. Cloning and Propagation:

22. 7. Characterization and Storage:

23. Production of Monoclonal Antibodies