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Mouse monoclonal antibodies (mAbs) play a pivotal role in both research and clinical applications. For research, they are crucial components in basic research and clinical laboratories, being indispensable in a variety of biological assays. Despite the traditional hybridoma generation and screening being a lengthy and low-throughput process, advancements such as genetic immunization, automation, and microarray applications have revolutionized the field, making the production of murine monoclonal antibodies faster and high-throughput.
The production of mouse monoclonal antibodies begins with injecting a mouse with an antigen to provoke an immune response. B-cells, a type of white blood cell, produce antibodies that bind to this antigen. These B-cells are then harvested and fused with immortal myeloma cancer cells to create a hybrid cell line known as a hybridoma. This fusion can be achieved using electrofusion or chemical methods. The resulting hybridomas are capable of both producing antibodies and replicating indefinitely. These hybridomas are then cultured, with each culture consisting of genetically identical cells producing monoclonal antibodies.
After fusion, the cells are incubated in HAT medium for about 10 to 14 days, allowing only the B cell-myeloma hybrids to survive and produce antibodies. The next steps involve primary screening processes like ELISA, immunocytochemical tests, and flow cytometry to select specific hybridomas that produce antibodies with the desired specificity. Once a desired hybridoma is identified, it can be cloned to produce many identical daughter clones. The antibodies produced are then harvested from the culture supernatant for subsequent investigations.
In clinical applications, mouse monoclonal antibodies have seen marked successes. Initially developed using hybridoma technology by Köhler and Milstein in 1975, these antibodies have transitioned from mouse, chimeric, humanized, to fully human forms, reducing potentially immunogenic mouse components. The US Food and Drug Administration has approved more than 20 mAbs, with over 150 others in clinical trials. However, mouse mAb therapy can be limited by the immunogenicity of the foreign protein, which can lead to adverse effects and loss of efficacy. Examples include Muromonab-CD3 causing cytokine release syndrome (CRS) and an acute influenza-like syndrome due to interaction with human anti-mouse antibodies. Despite these challenges, mAbs are established as targeted therapies for malignancies, transplant rejection, autoimmune and infectious diseases, among other indications.
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