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A multi‐dimensional approach for fractionating proteins using charged membranes
Author(s) -
Sorci Mirco,
Gu Minghao,
Heldt Caryn L.,
Grafeld Elizabeth,
Belfort Georges
Publication year - 2013
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.24837
Subject(s) - chemistry , ultrafiltration (renal) , membrane , lysozyme , chromatography , selectivity , ionic strength , filtration (mathematics) , analytical chemistry (journal) , biochemistry , organic chemistry , aqueous solution , catalysis , statistics , mathematics
In an effort to increase selectivity among proteins with crossflow ultrafiltration, we offer and demonstrate a comprehensive approach to fractionate proteins of similar molecular weight and relatively close p I values. This multidimensional approach involves optimizing membrane charge type and density together with operating conditions such as precise control of pH, ionic strength, and transmembrane pressure for reduced membrane fouling. Each filtration experiment was performed in cross‐flow configuration for ∼20 min, allowing fast screening for optimal separation as determined by maximum selectivity, Ψ , and purity, P . Using our comprehensive approach for fractionating mixtures RNase A–lysozyme and BSA–hemoglobin, we obtained values of Ψ = 9.1, P = 95.7%, and Ψ = 6.5, P = 62.1%, respectively. Biotechnol. Bioeng. 2013; 110: 1704–1713. © 2013 Wiley Periodicals, Inc.