Abstract
Epitope binning is a technique used to cluster different monoclonal antibodies by the specific region on the antigen that is recognized by the antibody. This is used to increase the likelihood of choosing lead antibodies with the desired biological activity by selecting antibodies from distinct bins, to confirm that biosimilars or biobetters bind to similar epitopes and to select reagents for diagnostic sandwich and/or ELISA-type assays. This protocol describes epitope binning of seven human monoclonal antibodies to a monovalent antigen in two orthogonal orientations; a classical sandwich and an in-tandem orientation using an eight-channel Octet system.
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References
Dong J, Demarest S, Sereno A et al (2010) Combination of two insulin-like growth factor-I receptor inhibitory antibodies targeting distinct epitopes leads to an enhanced antitumor response. Mol Cancer Ther 9:2593–2604
Koefoed K, Steinaa L, Soderberg L et al (2011) Rational indentiication of an optimal anitbody mixture for targeting the epidermal growth factor receptor. MAbs 3:584–595
Niederfellner G, Lammens A, Mudigl O et al (2011) Epitope characterization and chrsytal structure of GA101 provide insights into the molecular basis for type I/II distinction of CD20 antibodies. Blood 118:358–367
Byrd J, Kipps T, Flinn I et al (2012) Phase I study of the anti-CD40 humanized monoclonal anitbody lucatumumab (HCD122) in relapsed chronic lymphocytic leukemia. Leuk Lymphoma 53:2136–2142
Hellstrom K, Yelton D, Fell H et al (1990) Epitope mapping and use of anti-idiotypic antibodies to the L6 monoclonal anticarcinoma antibody. Cancer Res 50:2449–2454
Abdiche Y, Lindquist K, Stone D et al (2012) Label-free epitope binning assays of monoclonal antibodies enable identification of antigen heterogeneity. J Immunol Methods 382:101–116
Abdiche Y, Malashock D, Pinkerton A et al (2009) Exploring blocking assays using octet, ProteOn, and Biacore biosensors. Anal Biochem 386:172–180
Takkar R, Yadav A, Kumaraswamy S et al (2013) Cross-competition or epitope binning assays on the octet HTX system. Application Note of Pall Life Sciences, Menlo Park
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Bronswijk-Deddens, L. (2018). Epitope Binning of Human Monoclonal Antibodies in Classical Sandwich and In-Tandem Orientation Using the Octet System Based on Biolayer Interferometry. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_13
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DOI: https://doi.org/10.1007/978-1-4939-7841-0_13
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7839-7
Online ISBN: 978-1-4939-7841-0
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