Hypotonic elution, a new desorption principle in immunoadsorbent chromatography.
A largely unrecognized immunoadsorbent desorption technique, hypotonic elution, has been successfully used in the immunoadsorbent purification of the microvillar enzymes aminopeptidase N (EC 3.4.11.2), dipeptidyl peptidase IV (EC 3.4.14.5), sucrase-isomaltase (EC 3.2.1.48-10), lactase-phlorizin hydrolase (EC 3.2.1.23-62) and maltase-glucoamylase (EC 3.2.1.20). This elution method proved capable of achieving an acceptable yield (30-70%) while at the same time preserving the purified enzymes in an enzymically active state. It hereby offers a solution to the problem in immunoadsorbent chromatography of finding an efficient means of elution which is not denaturing to neither the purified enzyme nor the immunoadsorbent column. Common properties of the microvillar enzymes with regard to amphiphilicity, glycosylation or subunit composition could hypothetically account for the similar elution properties of the enzymes but were considered unlikely on several grounds. Hypotonic elution in immunoadsorbent chromatography, therefore, may have a much broader range of applicability, and the method is recommended to be tried out by workers in other areas of protein chemistry.
D000626:Aminopeptidases / Q000302:isolation & purification; D000818:Animals; D018826:CD13 Antigens; D002462:Cell Membrane / Q000201:enzymology*; D002845:Chromatography / Q000379:methods; D007038:Hypotonic Solutions; D007163:Immunosorbent Techniques; D008871:Microvilli / Q000201:enzymology*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011817:Rabbits; D013394:Sucrase-Isomaltase Complex / Q000302:isolation & purification; D013552:Swine; D014325:Tromethamine / Q000494:pharmacology*
D007038:Hypotonic Solutions; D014325:Tromethamine; D011092:Polyethylene Glycols; D017830:Octoxynol; D013394:Sucrase-Isomaltase Complex; D000626:Aminopeptidases; D018826:CD13 Antigens