hybridoma

Hybridoma is a hybrid cell line produced by fusing an antibody-producing B lymphocyte with a myeloma (cancer) cell, creating a cell that can both produce a specific antibody and proliferate indefinitely. Hybridomas are the basis for producing monoclonal antibodies (mAbs) with uniform specificity, essential for diagnostics and therapy.

Creation of hybridomas typically begins with immunizing a mouse with an antigen of interest, then isolating spleen B lymphocytes. These B cells are fused with immortal myeloma cells using a fusogenic agent such as polyethylene glycol. The fusion products are cultured in hypoxanthine-aminopterin-thymidine (HAT) medium; myeloma cells used are HGPRT-deficient and cannot survive in HAT, so only hybrids persist. Hybrids are then screened for the desired antibody specificity and cloned to ensure monoclonality.

Hybridoma lines are maintained in culture, and the antibodies are harvested from the supernatant or purified for use in diagnostics, research, and therapy. Monoclonal antibodies produced by hybridomas are valued for consistent binding characteristics and scalable production. Limitations include the murine origin of many early hybridomas, which can provoke immune responses in humans. This spurred the development of chimeric, humanized, and fully human monoclonal antibodies using recombinant DNA technologies, phage display, or transgenic animals.

Historically, Kohler and Milstein described the technique in 1975, revolutionizing biomedical research and medicine. Hybridomas are typically cultured under defined conditions, with antibodies purified by affinity chromatography. In modern practice, hybridoma genes may be cloned and expressed in recombinant systems, enabling alternative production hosts and humanized antibodies.