Abstract
Developments in the fields of bacterial expression of functional antibodies and methods to select genes from a library by using the phenotype of the encoded polypeptide have been a breakthrough in antibody technology. Today, phage display in combination with antibody gene libraries is widely used to select E. coli host cells that express desired antibody fragments. Such gene libraries are typically produced either from natural sources (e.g., from the spleen of an immunized animal or from plasma cells of human donors) or generated by genetic engineering. The latter has been used to create naïve libraries based on one or more antibody VH and VL gene segments that are diversified by cassette mutagenesis or similar approaches. Such libraries are typically unbiased and can be used for any given antigen (1–3). Modern naïve libraries are generally large (more than 1010 members), contain only few non-functional members, yield antibodies that are well expressed in E. coli (more than 1 mg of purified material per liter of culture) and are designed to allow further affinity maturation, if needed. Phage display is then most often used to select desired antibodies from such libraries (see refs. 4 and 5 for reviews).
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Knappik, A., Brundiers, R. (2009). Recombinant Antibody Expression and Purification. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-198-7_203
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DOI: https://doi.org/10.1007/978-1-59745-198-7_203
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