hybridoma technique

1. PROCEDURE OF HYBRIDOMA
TECHNOLOGY

2.  Lymphocytes are extracted from
spleen of an animal
 Immunized by antigen
 Subcutaneously or by peritoneal
injection

3.  Enhanced
production of B-
lymphocytes
 Increased chances of
required hybridoma
cells

4.  Antibody mixed with myeloma cells
 Not producing antibodies of their own
 Should contain genetic markers
hypoxanthine phospho-ribosyltransferase

5. Myeloma cells +
antibody producing lymphocytes
Hybrid cells + myeloma cells
+ B-lymphocytes

6.  Cultured in a selective media
 HAT medium alongwith drug
aminopterin
 Myeloma cells and B-lymphocytes will
die
 Resulting in hybrid cells
 Survival due to HGPRT from myeloma
cells
 Indefinite multiplication due to

7.  Various techniques
 ELISA (enzyme-linked immunosorbent assay)
 Hybrid cells producing the required
antibodies are isolated and cultured
 Ensures that single type of antibodies are
produced

8.  Cultured in suitable culture
media
 Injection of hybrid cells in
the peritoneal cavity
 Antibodies isolated from
the peritoneal fluid