Abstract
Antibodies are the fastest growing class of pharmaceutical proteins and essential tools for research and diagnostics. Often antibodies do show a desirable specificity profile but lack sufficient affinity for the desired application. Here, we describe a method to increase the affinity of recombinant antibody fragments based on the construction of mutagenized phage display libraries.
After the construction of a mutated antibody gene library by error-prone PCR, selection of high-affinity variants is either performed by panning in solution or on immobilized antigen with washing conditions optimized for off-rate-dependent selection. An additional screening protocol to identify antibodies with improved thermal stability is described.
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Unkauf, T., Hust, M., Frenzel, A. (2018). Antibody Affinity and Stability Maturation by Error-Prone PCR. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_22
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DOI: https://doi.org/10.1007/978-1-4939-7447-4_22
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7447-4
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