Peuravuori H, Kunelius R, Eskola J, Korpela T
Department of Pathology, University of Turku, Finland.
J Immunol Methods. 1997 May 26;204(2):161-7. doi: 10.1016/s0022-1759(97)00042-2.
beta 2-Microglobulin was purified from human peritoneal dialysate by ultrafiltration, gel chromatography and DEAE-high performance chromatography. Anti-beta 2-monoclonal antibodies were developed in mice and a pair of the antibodies was utilized to develop an enzyme-linked immunosorbent assay (ELISA) kit for the analyte with utilizing a new enzyme label, inorganic pyrophosphatase (EC 3.6.1.1). The sensitivity of the assay was 0.6 microgram/l (3 x SD) and the assay was linear up to absorbance values of around 2.0. No hook effect occurred in any putative concentrations of beta 2-microglobulin in serum. The precision of the assay of one run varied within 5-7% CV and the interassay precision was 2.8-8.6% CV, while the recovery was 99.2 +/- 6.0%. Excellent correlation occurred against an established radioimmunoassay method. All the used reagents were storable for a minimum of 1 year at +4 degrees C. It was decided that inorganic pyrophosphatase is a label of choice for ELISA.
通过超滤、凝胶色谱和DEAE高效色谱从人腹膜透析液中纯化β2-微球蛋白。在小鼠中制备抗β2-单克隆抗体，并利用一对抗体开发了一种酶联免疫吸附测定（ELISA）试剂盒，用于分析物的检测，该试剂盒使用了一种新的酶标记物——无机焦磷酸酶（EC 3.6.1.1）。该测定法的灵敏度为0.6微克/升（3×标准差），在吸光度值约为2.0之前测定呈线性。血清中任何假定浓度的β2-微球蛋白均未出现钩状效应。一次测定的精密度变异系数（CV）在5-7%以内，批间精密度CV为2.8-8.6%，回收率为99.2±6.0%。与既定的放射免疫测定方法具有良好的相关性。所有使用的试剂在4℃下至少可保存1年。确定无机焦磷酸酶是ELISA的首选标记物。
