Determination of a Small Amount of a Biological Constituent by the Use of Chemiluminescence. XII. Highly Sensitive Immunoaffinity Chromatography

Abstract

Abstract Highly sensitive immunoaffinity chromatography was established by combining a micro-bore affinity column with a chemiluminescence detector for protein monitored by a flow-injection method using a 1,10-phenanthroline–hydrogen peroxide–copper(II) system. After having optimized the bed volume of a column, the flow rate of a buffer solution which was used for an immune reaction, and the time necessary for an immune reaction in a column, human serum albumin in the range of 3×10−4–1×10−1 g dm−3 could be determined, with the detection limit of 15 ng as the injected amount. Human serum γ-globulin and bovine serum γ-globulin showed somewhat nonspecific adsorption onto an affinity column with values of 0.16 and 0.14 based on the specific adsorption 1.00 of human serum albumin onto a column, whereas bovine serum albumin, bovine serum α-globulin, and ovalbumin showed no nonspecific adsorption.