Abstract
High-affinity antibodies are crucial for development of monoclonal antibody (MAb)-based therapeutics for human diseases. Many new detailed methods for affinity maturation have been developed to improve MAb qualities by site-directed mutagenesis, chain shuffling, and error-prone PCR. Site-directed mutagenesis on hotspots in variable heavy (VH) complementary-determining region (CDR) 3 is a commonly used method for improving therapeutic potency and efficacy of targeted MAbs. Strategies for affinity maturation via multi-site-directed mutagenesis in VH-CDR3 described here are for valuable technical tool in the armamentarium of immunologists for development of fast-performance MAbs. Our strategy includes (1) selection of targeted MAb, (2) replacement of certain amino acid residues (e.g., negative or neutral charge to positive amino acids) in VH-CDR3, and (3) determination of binding activity to a target antigen.
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Acknowledgment
We thank Tommy Kim for editorial assistance.
Disclaimer. The views expressed in this manuscript are those of the authors and do not reflect the official policy of the Department of the Army, the Department of Defense, the US Government, or the Henry M Jackson Foundation for the Advancement of Military Medicine.
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Kim, HY., Stojadinovic, A., Izadjoo, M.J. (2014). Affinity Maturation of Monoclonal Antibodies by Multi-Site-Directed Mutagenesis. In: Ossipow, V., Fischer, N. (eds) Monoclonal Antibodies. Methods in Molecular Biology, vol 1131. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-992-5_24
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DOI: https://doi.org/10.1007/978-1-62703-992-5_24
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