犬心脏组织组织蛋白酶D的双抗体放射免疫测定法。

A double antibody radioimmunoassay for canine cardiac cathepsin D.

作者信息

Samarel A M, Ogunro E A, Ferguson A G, Lesch M

出版信息

Cardiovasc Res. 1981 Feb;15(2):68-73. doi: 10.1093/cvr/15.2.68.

DOI:10.1093/cvr/15.2.68

Abstract

We have developed a sensitive double antibody radioimmunoassay for measuring canine cardiac cathepsin D. Radioiodinated cathepsin D was prepared by chloramine T oxidation using a highly purified source of enzyme. High avidity antiserum to the canine cardiac enzyme was raised in rabbits. Antibody-bound cathepsin D was separated from free enzyme using goat anti-rabbit IgG second antibody. The least amount of immunoreactive enzyme measurable in the radioimmunoassay was 2.4 ng.cm-3 as determined by antibody titration. The assay was linear for concentrations of enzyme in the range of 10 to 120 ng.cm-3. Within-assay and between-assay variations were 12%. The radioimmunoassay described was used to measure the immunoreactive cathepsin D content of the 100 000 x g supernatant fraction of canine myocardial homogenates.

摘要

我们开发了一种用于测量犬心脏组织组织蛋白酶D的灵敏双抗体放射免疫分析方法。使用高度纯化的酶源，通过氯胺T氧化法制备放射性碘化组织蛋白酶D。在兔体内产生了对犬心脏酶的高亲和力抗血清。使用山羊抗兔IgG二抗将抗体结合的组织蛋白酶D与游离酶分离。通过抗体滴定法测定，放射免疫分析中可测量的最低免疫反应性酶量为2.4 ng·cm⁻³。该分析方法在酶浓度为10至120 ng·cm⁻³范围内呈线性。批内和批间变异为12%。所描述的放射免疫分析方法用于测量犬心肌匀浆100 000×g上清液部分的免疫反应性组织蛋白酶D含量。