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Structural and functional characteristics of virgin and fouled Protein A MabSelect resin cycled in a monoclonal antibody purification process
Author(s) -
Zhang Shaojie,
Xu Kerui,
Daniels William,
Salm Jeffrey,
Glynn Judy,
Martin Joseph,
Gallo Christopher,
Godavarti Ranga,
Carta Giorgio
Publication year - 2016
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.25708
Subject(s) - chromatography , adsorption , desorption , chemistry , size exclusion chromatography , porosity , kinetics , scanning electron microscope , chemical engineering , materials science , analytical chemistry (journal) , composite material , biochemistry , organic chemistry , physics , quantum mechanics , engineering , enzyme
The structural and functional characteristics of the Protein A MabSelect resin are determined for a virgin sample and for samples removed from a column that had been operated in an antibody capture process which had shown losses in product recovery over fewer than 20 cycles. Compared to the virgin resin, the cycled samples show reduced porosity and apparent pore size based on inverse size exclusion chromatography while transmission electron microscopy (TEM) shows accumulation of foulants on the cycled resin. Adsorption isotherms, batch adsorption kinetics, and batch desorption kinetics, obtained using the antibody in purified form, show that the cycled samples have about 10% lower binding capacity and slower mass transfer. Confocal scanning laser microscopy shows, however, that different degrees of fouling exist for different beads in the cycled samples, which may correspond to the existence of areas exposed to minimal or no flow in the process column. Replacing the standard cleaning procedure with an improved multi‐step cleaning protocol prevented the accumulation of foulants in the resin beads, as evident from TEM, and resulted in a stable operation with high recovery. Biotechnol. Bioeng. 2016;113: 367–375. © 2015 Wiley Periodicals, Inc.