Abstract
The ion exchange-high-performance liquid chromatography is a high-throughput analytical method that allows to determine the charge profile of purified antibodies. Here, we describe the preparation of the samples, chromatographic conditions to be used (buffer preparation, salt gradient, quantity injected, flow rate, run time, column suitability, etc.), validity of the analysis, and integration of the chromatogram in order to calculate the proportion of the different isoforms.
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References
Melter L, Ströhlein G, Butté A, Morbidelli M (2007) Adsorption of monoclonal antibody variants on analytical cation-exchange resin. J Chromatogr A 1154:121–131
Macchi FD, Shen FJ, Kwong M, Andya JD, Shire SJ, Bjork N, Totpal K, Chen AB (2001) Identification of multiple sources of charge heterogeneity in a recombinant antibody. J Chromatogr B Biomed Sci Appl 752:233–245
Vlasak J, Ionescu R (2008) Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. Curr Pharm Biotechnol 9:468–481
Rea JC, Moreno GT, Lou Y, Farnan D (2011) Validation of a pH gradient-based ion-exchange chromatography method for high-resolution monoclonal antibody charge variant separations. J Pharm Biomed Anal 54(2):317–323
Zhang T, Bourret J, Cano T (2011) Isolation and characterization of therapeutic antibody charge variants using cation exchange displacement chromatography. J Chromatogr A 1218(31):5079–5086
Liu H, Gaza-Bulseco G, Faldu D, Chumsae C, Sun J (2008) Heterogeneity of monoclonal antibodies. J Pharm Sci 97(7):2426–2447
Rozhkova A (2009) Quantitative analysis of monoclonal antibodies by cation-exchange chromatofocusing. J Chromatogr A 1216(32):5989–5994
Khawli LA, Goswami S, Hutchinson R, Kwong ZW, Yang J, Wang X, Yao Z, Sreedhara A, Cano T, Tesar D, Nijem I, Allison DE, Wong PY, Kao Y-H, Quan C, Joshi A, Harris RJ, Motchnik P (2010) Charge variants in IgG1. mAbs 2(6):613–624
402 LCGC NORTH AMERICA (2006) 24(4) www.chromatographyonlime.com
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Corbier, M., Schrag, D., Raimondi, S. (2014). Ion Exchange-High-Performance Liquid Chromatography (IEX-HPLC). 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_30
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DOI: https://doi.org/10.1007/978-1-62703-992-5_30
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-991-8
Online ISBN: 978-1-62703-992-5
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