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-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).
Kasche
V
V
AB Biotechnologie II, Technical University of Hamburg-Harburg, F.R.G.
Löffler
F
F
Scholzen
T
T
Krämer
D M
DM
Boller
T
T
D000327:Adsorption; D002082:Butylamines; D055598:Chemical Phenomena; D002621:Chemistry; D004926:Escherichia coli / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010399:Penicillin Amidase / Q000302:isolation & purification; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D002082:Butylamines; D011506:Proteins; D012997:Solvents; C065841:phenylbutylamine-Eupergit; D010399:Penicillin Amidase