Johansen L, Nustad K, Orstavik T B, Ugelstad J, Berge A, Ellingsen T
J Immunol Methods. 1983 Apr 29;59(2):255-64. doi: 10.1016/0022-1759(83)90038-8.
The development of an excess antibody assay for rat glandular kallikrein is described. This assay permits immunological determination of kallikrein as well as a simultaneous specific measurement of kallikrein enzymatic activity. The assay is based on coupling of immunopurified anti-kallikrein immunoglobulin to a solid phase. In a first incubation step, kallikrein was bound to the immobilized antibody. Determination of kallikrein was subsequently done in a second incubation step; immunologically by addition of iodinated anti-kallikrein antibody, or enzymatically by a kallikrein substrate. Enzymatic quantification could also be followed by immunological measurements on the same sample. Comparison of Sepharose, cellulose, and acrylate based polymer particles proved the latter to be the best matrix in this assay. The main advantage of the polymer particles was the low non-specific binding of labelled antibody.
本文描述了一种用于大鼠腺体激肽释放酶的过量抗体检测方法的开发。该检测方法可通过免疫学方法测定激肽释放酶，并同时特异性测量激肽释放酶的酶活性。该检测方法基于将免疫纯化的抗激肽释放酶免疫球蛋白偶联到固相上。在第一步孵育中，激肽释放酶与固定化抗体结合。随后在第二步孵育中进行激肽释放酶的测定；通过加入碘化抗激肽释放酶抗体进行免疫学测定，或通过激肽释放酶底物进行酶学测定。酶学定量之后也可以对同一样品进行免疫学测量。对基于琼脂糖、纤维素和丙烯酸酯的聚合物颗粒进行比较，结果证明后者是该检测方法中最佳的基质。聚合物颗粒的主要优点是标记抗体的非特异性结合较低。
