Bahar I
Hygeia Sciences, Newton, MA 02160.
Prog Clin Biol Res. 1988;285:139-51.
The development of simple tests for estrone-B,D-glucuronide (E1G) and pregnanediol-3-glucuronide (P3G) in urine is described. The haptens P3G and E1G, coupled to bovine serum albumin (BSA), were used as immunogens against which specific monoclonal antibodies were made by fusion of variants of P3.X63.Ag8.653 with spleen lymphocytes from immunized mice. When covalently bonded to gelatin or BSA and passively adsorbed to a microtiter plate, the hapten provides the solid phase for an ELISA. Peroxidase-labelled monoclonal antibody is premixed with a urine specimen and the mixture is immediately added to the plate. After a brief incubation and washing, a mixture of tetramethylbenzidine chromogen and hydrogen peroxide is added to serve as substrate and generate color. The ELISA can be used to monitor levels of E1G and P3G during menstrual cycles and provides a simple, noninvasive method which can be used in a laboratory. A similar competitive assay can be performed using colloidal gold as the label instead of peroxidase. The replacement of peroxidase by colloidal gold further simplifies the procedure and could be used as the basis for an OTC test.
本文描述了尿液中雌酮 - B，D - 葡萄糖醛酸苷（E1G）和孕二醇 - 3 - 葡萄糖醛酸苷（P3G）简单检测方法的开发。将与牛血清白蛋白（BSA）偶联的半抗原P3G和E1G用作免疫原，通过P3.X63.Ag8.653变体与免疫小鼠的脾淋巴细胞融合制备特异性单克隆抗体。当半抗原共价结合到明胶或BSA上并被动吸附到微量滴定板上时，它为酶联免疫吸附测定（ELISA）提供固相。过氧化物酶标记的单克隆抗体与尿液标本预混合，然后立即将混合物加入板中。短暂孵育和洗涤后，加入四甲基联苯胺色原和过氧化氢的混合物作为底物显色。该ELISA可用于监测月经周期中E1G和P3G的水平，并提供一种可在实验室中使用的简单、非侵入性方法。也可以使用胶体金代替过氧化物酶作为标记进行类似的竞争性测定。用胶体金替代过氧化物酶进一步简化了操作程序，可作为非处方测试的基础。
