Hybridoma Technology for the Generation of Monoclonal Antibodies
Hybridoma technology has long been a remarkable and indispensable platform for generating high-quality monoclonal antibodies (mAbs). Hybridoma-derived mAbs have not only served as powerful tool reagents but also have emerged as the most rapidly expanding class of therapeutic biologics. With the establishment of mAb humanization and with the development of transgenic-humanized mice, hybridoma technology has opened new avenues for effectively generating humanized or fully human mAbs as therapeutics. In this chapter, an overview of hybridoma technology and the laboratory procedures used routinely for hybridoma generation are discussed and detailed in the following sections: cell fusion for hybridoma generation, antibody screening and characterization, hybridoma subcloning and mAb isotyping, as well as production of mAbs from hybridoma cells.
Key wordsCell fusion ELISA Flow cytometry Hybridoma technology Immunohistochemistry Immunization Monoclonal antibody Myeloma cells Therapeutic antibody Screening
The author gratefully acknowledges Lena Kikuchi and Nancy Chan for their expertise in hybridoma generation and the technical assistance for validating the protocols described here. The author is especially thankful to Dr. Peter LeMotte, Dr. Thomas Pietzonka and Dr. John Hastewell for helpful advice and support, and to Yuxiang Zhang for his assistance in editing the illustrations in this manuscript.
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