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Enzyme separation by parametric pumping
Author(s) -
Shaffer A. G.,
Hamrin C. E.
Publication year - 1975
Publication title -
aiche journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.958
H-Index - 167
eISSN - 1547-5905
pISSN - 0001-1541
DOI - 10.1002/aic.690210421
Subject(s) - trypsin , chemistry , chromatography , elution , aqueous solution , chymotrypsin , covalent bond , sepharose , parametric statistics , affinity chromatography , analytical chemistry (journal) , enzyme , biochemistry , organic chemistry , mathematics , statistics
Abstract Affinity chromatography and parametric pumping were combined to reduce trypsin concentration in an aqueous solution. The trypsin inhibitor, chicken ovomucoid, was covalently bonded to Sepharose 4B beads. A solution was cycled over this packing in a column at a low p H in one direction followed by a high p H in the other direction. Trypsin retention was favored at the high p H and elution at the low p H. The decrease in trypsin concentration at one end of the column was fitted to an equation derived from the equilibrium parametric pumping model and was a function of the p H limits imposed on the column. Separation was much less than predicted by equilibrium data, but the equation based on trypsin reduction agreed with the α‐chymotrypsin‐trypsin separation.