IMPROVED ION-EXCHANGED HPLC METHOD IN mAb USING pH GRADIENT AND ITS COMPARISON WITH cIEF

Autor:	Carrie L. Anderson, Yang Wang, Brian Peklansky, Richard R. Rustandi, Feng Wang
Rok vydání:	2012
Předmět:	Chromatography Resolution (mass spectrometry) Isoelectric focusing medicine.drug_class Chemistry Clinical Biochemistry Ion chromatography Analytical chemistry Pharmaceutical Science Fraction (chemistry) Monoclonal antibody Biochemistry Analytical Chemistry Ion Ph gradient medicine Hplc method
Zdroj:	Journal of Liquid Chromatography & Related Technologies. 35:1259-1269
ISSN:	1520-572X 1082-6076
DOI:	10.1080/10826076.2011.619035
Popis:	Ion-exchange chromatography (IEX) and capillary isoelectric focusing (cIEF) are two common methods used for molecular charge heterogeneity analysis of therapeutic proteins. The two methods are complementary to each other and each method has its own advantage and disadvantages. For example, a sample injected on an IEX column can be easily fraction collected but, sometimes, it lacks the ability to provide good separation of charge variants, such as those present in monoclonal antibody (mAb) samples. While cIEF has much better resolution than IEX, the separated peaks cannot be collected. Recently, pIsep, a new ion chromatographic technique developed by CryoBioPhysica, has been successfully applied to analyze charged heterogeneities in mAbs. The pIsep separation utilizes an external pH gradient and the charged variants are resolved based on pI differences. In this study, an acidic variant co-eluted with main peak on traditional salt-gradient cation-exchange chromatography (CEX) is separated by pIsep and the f...
Databáze:	OpenAIRE
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