胰岛素放射免疫分析质控血清的研制

PREPARATION OF INSULIN QUALITY CONTROL SERA FOR RADIOIMMUNOASSAY

介绍了亲和层析法制备胰岛素零值血清和胰岛素放射免疫分析质控血清研制的过程。胰岛素为免疫原对豚鼠进行免疫获得抗血清，将此抗血清直接偶联到溴化氰活化Ｓｅｐｈａｒｏｓｅ４Ｂ凝胶上，制成亲和层析柱，使正常人血清通过此柱，胰岛素被特异地吸附在柱中而从血清中除掉。用放免方法测定过柱处理后的血清，其中不含胰岛素，经蛋白测定和醋酸纤维素膜电泳检测证明过柱前、后血清蛋白含量和组成未产生明显变化。向此血清中定量加入胰岛素并经标定确定其靶值和允许范围，即制成低、中和高值质控血清。

Anti insulin sera are obtained from male guinea pigs immunized with porcine insulin and are used for preparation of affinity chromatographic column. The column is packed with CNBr activated Sepharose 4B coupling with anti insulin sera. When normal human sera are passing through the affinity chromatographic column, insulin is adsorbed on the column. After passed through the column, these sera are insulin free by RIA and protein composition of the sera is similar to that before the treatment by cellulose acetate of membrane electropharesis. The insulin quality control (QC) sera is prepared using insulin free sera in which human insulin added quantitativelly. The QC sera (low, medium and high concentration) are calibrated, and the reference values and their limits are obtained from statistical results of calibration.