Comparison of low- and high-pressure affinity chromatography for the purification of serine and sulfhydryl esterases.
A series of affinity chromatography packings for the purification of serine and sulfhydryl esterases (acetylcholinesterase, alkaline phosphatase, urokinase and papain) have been synthesized using commercially available agarose, glass and acrylate parent matrices. Two ligands were coupled to the matrices by utilizing carbodiimide or reaction with active groups already present on the matrix: the quaternary ammonium compound trimethyl(p-aminophenyl)ammonium chloride and the serine esterase inhibitor analog p-aminobenzamidine. It was found that enzyme purification on the agarose- or acrylate-based packings was most successful, resulting in as much as fifty-fold purification over starting material. The pressure stability of the acrylate packing allowed purification by high-pressure affinity chromatography and decreased purification times as much as six-fold in comparison to agarose columns.
D000110:Acetylcholinesterase / Q000302:isolation & purification; D000469:Alkaline Phosphatase / Q000302:isolation & purification; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004557:Electric Organ / Q000201:enzymology; D004593:Electrophorus; D004950:Esterases / Q000302:isolation & purification*; D010206:Papain / Q000302:isolation & purification; D011485:Protein Binding; D014357:Trypsin / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000302:isolation & purification
D004950:Esterases; C022635:serine esterase; D000110:Acetylcholinesterase; D000469:Alkaline Phosphatase; D014357:Trypsin; D014568:Urokinase-Type Plasminogen Activator; D010206:Papain