Hybridoma Technology: Process, Applications & Monoclonal Antibody Production

                                   Hybridoma Technology 

Hybridoma Technology is a method used to produce hybrid cells by fusing a specific antibody-producing B-cell with a myeloma (cancer) cell. The process begins by injecting a specific antigen into a mouse to stimulate an immune response. The antibody-producing B-cells are then collected from the mouse’s spleen and fused with myeloma cells.

This fusion results in hybrid cells, known as hybridomas, which have two key properties: the ability to produce a specific antibody (from the B-cell) and the ability to divide indefinitely (from the myeloma cell).

The fused cells are then placed in a special culture medium where the unfused myeloma cells die, but the hybridomas survive. This is achieved by using HAT medium (which contains hypoxanthine, aminopterin, and thymidine).

Aminopterin in the medium blocks the de novo nucleotide synthesis pathway, so cells must use the salvage pathway to survive. However, myeloma cells used in the fusion are HGPRT-deficient (they lack hypoxanthine-guanine phosphoribosyltransferase), meaning they cannot use the salvage pathway and die in the presence of aminopterin.

Only the hybridomas, which inherit the HGPRT enzyme from the B-cells, can survive in HAT medium by using the salvage pathway. These surviving hybridoma cells are then cultured and screened to identify those producing the desired antibody. In this way, monoclonal antibodies are generated.