Describe the technology of making monoclonal antibodies and

Describe the technology of making “monoclonal antibodies”, and be able to explain how HAT media works to select for hybridoma cells. Explain some pros vs cons of using monoclonal antibodies.

Solution

Answer:

The B lymphocytes are isolated from the spleen of an animal that has been challenged with the relevant antigen and fused with myeloma tumor cells. The fused cells -called hybridomas will multiply indefinitely to produce large amounts of the desired antibodies.

Fused cells are incubated in the medium containing Hypoxanthine Aminopetrin Thymidine (HAT). Aminopterin in the media blocks nucleotide synthesis. Hence, unfused myeloma cells die, as they cannot produce nucleotides by the de novo or salvage pathways because they lack hypoxanthine-guanine phosphoribosyltransferase. Unfused B cells die as they have a short life span