Recklies A D, Mort J S
Clin Chim Acta. 1982 Aug 4;123(1-2):127-38. doi: 10.1016/0009-8981(82)90121-8.
A radioimmunoassay for human cathepsin B is described, which demonstrates the usefulness of protein A-bearing Staphylococcus aureus as an immunosorbent in a system where the primary antibody IgG has a low binding affinity for protein A. The removal of bound antigen from the incubation mixture is achieved by the use of a rabbit second step antiserum which confers high binding affinity for protein A to the primary immune complex. This method, as employed in the assay for human cathepsin B, is very reproducible and economical for large numbers of samples. The use of a monospecific antiserum to human cathepsin B and slightly alkaline assay conditions allow the determination of total cathepsin B protein in tissue fluids which is not possible by enzyme activity determination.
本文描述了一种人组织蛋白酶B的放射免疫测定法,该方法证明了携带蛋白A的金黄色葡萄球菌作为免疫吸附剂在一系统中的实用性,在此系统中,一抗IgG对蛋白A的结合亲和力较低。通过使用兔二抗血清从孵育混合物中去除结合抗原,该二抗血清赋予初级免疫复合物对蛋白A高结合亲和力。在人组织蛋白酶B的测定中采用的此方法,对于大量样本而言具有高度可重复性且经济实用。使用人组织蛋白酶B的单特异性抗血清以及略呈碱性的测定条件,能够测定组织液中的总组织蛋白酶B蛋白,而通过酶活性测定则无法做到这一点。