Rapid protein purification using phenylbutylamine-Eupergit: a novel method for large-scale procedures

Electrophoretic desorption was used to compare the protein binding capacities of some hydrophobic adsorbents [the phenylbutylamine (PBA) derivatives of Eupergit C and agarose and Phenyl-Sepharose] for low-pressure chromatography. The highest capacity was observed for the bifunctional adsorbent PBA-E...

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Bibliographic Details
Published inJournal of chromatography Vol. 510; p. 149
Main Authors Kasche, V, Löffler, F, Scholzen, T, Krämer, D M, Boller, T
Format Journal Article
LanguageEnglish
Published Netherlands 27.06.1990
Subjects
Adsorption
Butylamines
Chemical Phenomena
Chemistry
Escherichia coli - enzymology
Hydrogen-Ion Concentration
Kinetics
Penicillin Amidase - isolation & purification
Protein Binding
Proteins - isolation & purification
Solvents
Spectrophotometry, Ultraviolet
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Summary:Electrophoretic desorption was used to compare the protein binding capacities of some hydrophobic adsorbents [the phenylbutylamine (PBA) derivatives of Eupergit C and agarose and Phenyl-Sepharose] for low-pressure chromatography. The highest capacity was observed for the bifunctional adsorbent PBA-Eupergit. The hydrophobically adsorbed proteins can be selectively desorbed by decreasing the pH of the eluent due to electrostatic repulsion between positive charges on the adsorbed proteins and positively charged secondary amines on the adsorbent. This was used to purify 1500 U penicillin amidase from E. coli homogenates per gram wet weight of PBA-Eupergit in 50 adsorption-desorption cycles without organic solvents (greater than 90% yield, purification factor = 5.3).
DOI:10.1016/S0021-9673(01)93748-3