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Displacement effect during HPLC preparative purification of human insulin.

作者信息

Gusarov D, Nekipelova V, Gusarova V, Lasman V, Bairamashvili D

机构信息

Experimental Biotechnological Plant, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Moscow, Russian Federation.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 15;877(11-12):1216-20. doi: 10.1016/j.jchromb.2009.02.062. Epub 2009 Mar 5.

DOI:10.1016/j.jchromb.2009.02.062
PMID:19286429
Abstract

HPLC plays a key role in the preparative purification of human insulin. A21-desamidoinsulin is one of the impurities that possesses the chromatographic behavior similar to that of insulin and hence separation from this by-product is rather difficult at the process scale. During the optimization of insulin reversed-phase HPLC purification, when a column was sufficiently overloaded, the effect of displacement of A21-desamidoinsulin molecules from active groups of sorbent by insulin ones was observed. It was suggested that monocarboxylic acid and organic modifier in mobile phase are responsible for the esterification during which the formed ester promotes the displacement effect. This effect was studied in order to optimize the purification of human insulin at the process scale.

摘要

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