Hybridomas are the result of a fusion between an antibody producing spleen cell and an immortal myeloma cell.
How Hybridomas are Created
Immunization and B Lymphocyte Isolation
Hybridoma production begins by immunizing laboratory animals, typically mice, with the antigen of interest. This antigen stimulates the mouse’s immune system to produce B lymphocytes that generate antibodies targeting the specific antigen. After a series of immunizations, the spleen—rich in activated B cells—is removed for isolation of the B lymphocytes that now carry genetic information to produce antigen-specific antibodies.
Fusion of B Cells with Myeloma Cells
Isolated B lymphocytes are then fused with immortalized myeloma cells, which are cancerous plasma cells with a high proliferation capacity. This fusion process uses polyethylene glycol (PEG) to promote the merging of cell membranes, creating hybrid cells or "hybridomas." The myeloma cells provide longevity, while the B lymphocytes contribute the antibody-producing capacity. The hybridoma cells that form are effectively immortal, allowing for indefinite production of a consistent, highly specific monoclonal antibody.
Selection and Screening
Hybridoma cells are cultured in a selective medium (HAT medium) to filter out unfused cells. Only hybridomas—those with both B cell and myeloma cell properties—survive, as they possess both the antibody-coding genes from B cells and the longevity from myeloma cells. Once selected, the hybridomas are screened to identify those producing the desired antibody, which is subsequently purified for applications in research and therapeutics.
Advantages of Hybridoma Technology
Hybridoma-derived antibodies are highly specific
Reproducible and scalable, ensuring consistency across large batches
Unlimited production of monoclonal antibodies
Useful for highly sensitive and specific assays
Purity of antigen or immunogen is not a prerequisite
Not labor-intensive as in vitro antibody generation techniques uses immune libraries
Once the hybridoma line is established, the cost per antibody unit decreases
Advances in genome-editing tools, like CRISPR, offer the potential to improve the precision of human antibody production, reducing the need for traditional animal-derived hybridomas and potentially more diverse and human-compatible mAb therapies. As hybridoma technology continues to evolve, we can expect to see further improvements in monoclonal antibody production efficiency, specificity, and safety—transforming hybridoma technology from a classic method to a modern powerhouse in therapeutic antibody production.
References:
Mitra, S., & Tomar, P. C. (2021). Hybridoma technology; advancements, clinical significance, and future aspects. Journal of Genetic Engineering & Biotechnology, 19, 159. https://doi.org/10.1186/s43141-021-00264-6
