In 1975, Köhler and Milstein discovered the hybridoma technique to generate monoclonal antibodies. Hybridoma cells are formed by fusing a short-lived antibody-producing B cell with an immortal myeloma cell. The immunized animal-derived spleen cells are the source of B cells and the mouse/rat-derived myeloma cell lines are used as fusion parents. In vitro experiments, these two cells have the properties necessary for a successful hybridoma cell. The B cells will provide the functional immunoglobulin genes specific to one antigenic epitope, and the myeloma cells possess growth properties, rapidly dividing and lacking the salvage pathway of DNA synthesis. Finally, hybridoma cells could secrete monoclonal antibodies and could be established routinely.
Figure 1, 2 In vitro experiments to scale up and culture the hybridoma cells.
At present, hybridoma technology remains the primary discovery platform preferred by in vivo scientists. The hybridoma cells-derived monoclonal antibodies not only provide a basic understanding of science but also an advantage in vivo research, diagnostic tests, and therapy. The application of hybridoma cells-derived monoclonal antibodies is widespread in humans. For example, the use of monoclonal antibodies in therapy for emerging and re-emerging infectious diseases i.e., Rabies, Ebola, and COVID-19. The use of monoclonal antibodies in diagnostic tests against various pathogens, including viruses, bacteria (mainly intracellular), fungi, and parasites.
In contrast to humans, the application of monoclonal antibodies in animals mainly focused on diagnostic tests rather than prophylaxis or treatment. In animals, the use of monoclonal antibodies to detect antigens and antibodies of microbial diseases by using different techniques such as western blotting, enzyme-linked immunosorbent assay, immunofluorescence test, immunohistochemistry, and immunochromatography. For example, bacterial diseases (i.e., anthrax, brucellosis, paratuberculosis, leptospirosis, clostridial diseases, etc.); fungal diseases (i.e., aspergillosis, candidiasis, cryptococcosis, histoplasmosis, etc.); viral diseases (i.e., foot-and-mouth disease, rabies, Hendra virus, Nipah virus, etc.); and parasitic diseases (i.e., trichinellosis, trypanosomiasis, leishmaniasis, etc.) have been demonstrated. In addition, the further development of monoclonal antibodies to identify novel biomarkers may be helpful for antibody-based therapy for animal diseases.
Reference
Kohler G and Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975, 256, 495-497.
Deb R, Chakraborty S, Veeregowda B, Verma AK, Tiwari R, Dhama K. Monoclonal antibody and its use in the diagnosis of livestock diseases. Adv. Biosci. Biotechnol. 2013, 4, 50-62.
Holzlöhner P and Hanack K. Generation of Murine Monoclonal Antibodies by Hybridoma Technology. J. Vis. Exp. 2017, 119, 54832.
Weena Paungpin
Researcher, The Monitoring and Surveillance Center for Zoonotic Diseases in Wildlife and Exotic Animals (MoZWE)