In recent years, antibodies have become increasingly accepted as therapeutics for human diseases, particularly for cancer, viral infection and autoimmune disorders. Monoclonal antibodies (Mabs) have been used as diagnostic and analytical reagents since hybridoma technology was invented in 1975. “man-made antibodies.” was named by Cesar Milstein, who was one of the inventors of monoclonal antibody technology. Until the late 1980’s, antibody technology relied primarily on animal immunization and the expression of engineered antibodies.

1. ANTIBODY
ENGINEERING
ARCHANA GAUTAM
M.Sc. 3rd
Sem.
BBAU lucknow

2. ANTIBODY ENGINEERING
• ANTIBODIES
• CHIMERIC /HUMANIZED
• METHODOLOGIES
• APPLICATIONS

3. Cont....
• In recent years, antibodies have become increasingly
accepted as therapeutics for human diseases, particularly
for cancer, viral infection and autoimmune disorders.
• Monoclonal antibodies (Mabs) have been used as diagnostic
and analytical reagents since hybridoma technology was
invented in 1975.
• “man-made antibodies.” was named by Cesar Milstein, who
was one of the inventors of monoclonal antibody
technology.
• Until the late 1980’s, antibody technology relied primarily
on animal immunization and the expression of engineered
antibodies.

4. PROBLEMS
• HAMA Response.
• Loss of effector
functions.
• Reduce the size of
antibody.

5. LIGHT CHAIN GENES
Variable
domain
CDR3

6. HEAVY CHAIN GENES
HUMAN
H.C Variable region= V+D+J gene segments

9. • Production of
human Mab.
• Make mouse Ab
more “human-like”
• Production of small
Ab fragments.
SOLUTIONS

10. Human Mouse
Chimeric

11. HUMANIZED ANTIBODIES
FR1 FR2 FR3 FR4
CDR1 CDR2 CDR3
CONSTANT
CONSTANT
CONSTANT
FR1 FR2 FR3 FR4
CDR1 CDR2 CDR3
Mouse
Human
Humanized

12. Human Mouse
Humanized

13. CHIMERIC/HUMANIZED
.Reduce the
HAMA response.
.Maintain effector
functions.
.Increase the half
life of the antibody.
.Maintain binding
affinity.
.Easy to construct.

14. ANTIBODY
ENGINEERING

15. .HAMA Response.
•Loss of effector
functions.
Making the
antibody more
“human-like”. (the
cutting and pasting)
Production of
human Mab’s.
Chimeric Ab’s Humanized Ab’s
Hybridoma
Phage Display
Transgenic/SCID
Mice

16. MONOCLONAL ANTIBODIES
Mouse
splenocytes
Mouse
myeloma
hybridoma
Y Y YMonoclonal
Y Y
Antibodies
Y Y

17. HUMAN HYBRIDOMAS
. Unstable
hybridomas.
.Small number of
human B-cells in
peripheral blood.
. Obtaining
antigen-reactive
human B-cells.
Heteromyeloma
fusion
In vitro
immunization

18. MICE
SCID
Transgenic
Reconstituted
with human
peripheral blood
Containing
human
immunoglobulin
genes.

19. PHAGE DISPLAY

20. Reduce the
size of
Antibody. Production of
antibody
fragments.
Simplifying the
expression/production
of antibodies.

21. ANTIBODY FRAGMENTS

22. EXPRESSION SYSTEMS
FungalMammalian cell
lines
Bacterial

23. Application

24. Thank u