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Size‐exclusion reaction chromatography (SERC): A new technique for protein PEGylation
Author(s) -
Fee Conan J.
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10561
Subject(s) - chemistry , pegylation , chromatography , size exclusion chromatography , yield (engineering) , residence time (fluid dynamics) , separation process , peg ratio , materials science , organic chemistry , polyethylene glycol , geotechnical engineering , finance , economics , engineering , metallurgy , enzyme
A new, widely applicable process that combines reaction and separation in a single unit operation is described. The process, size‐exclusion reaction chromatography (SERC), simultaneously allows control of the extent of reactions in which molecular size is altered and the separation of products and reactants. In SERC, a moving reaction zone is formed by injection of reactants onto a size‐exclusion chromatography column. Reactants and products are partitioned differently within the mobile phase, resulting in different linear flow rates through the column. The products are therefore removed selectively from the reaction zone, minimizing their residence time in the reaction zone and allowing their separation in the downstream section of the column. For reactions such as protein PEGylation, in which successive addition of PEG groups to the protein results in significant molecular size increases, SERC potentially offers a method by which a dominant final PEGylated protein size can be produced at high yield. The SERC PEGylation of two model proteins, α‐lactalbumin and β‐lactoglobulin, is demonstrated and results show that simultaneous reaction and separation was obtained. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 200–206, 2003.