Chu T M, Kuriyama M, Johnson E, Papsidero L D, Killian C S, Murphy G P, Wang M C
Ann N Y Acad Sci. 1983;417:383-9. doi: 10.1111/j.1749-6632.1983.tb32880.x.
A reverse enzyme-linked immunosorbent assay (ELISA) modified from a prostate antigen (PA) assay previously reported has been developed to measure circulating PA-binding globulin (PABG). Serum specimens taken from normal males, normal females, male patients with nonprostatic cancers, patients with benign prostatic hypertrophy, and patients with various stages of prostate cancer were analyzed for PABG. Results revealed that only patients with an advanced stage of prostatic cancer exhibited an elevated level of PABG. PABG was then isolated from prostatic cancer patients' serum by PA affinity chromatography. Upon immunodiffusion and immunoelectrophoresis, this PABG preparation reacted with purified PA, anti-PA xenoantibodies and anti-human IgG. By the immunoperoxidase technique, PABG stained positively in prostatic ductal epithelial cells and negatively in all other tissues examined. An additional PABG preparation, which reacted with anti-PA and anti-human IgG and not with purified PA, was also isolated by an anti-PA affinity chromatography. These PABG preparations were separately subjected to high-performance liquid chromatography for further purification. Three major protein peaks at Mr of greater than 240K, 150K, and 34K were obtained. These results demonstrated that circulating IgG antibody reactive with PA was present in patients with metastatic cancer of the prostate, and this in part was in complexed form with PA and was specifically reactive with ductal epithelial elements of the prostate.
我们改进了先前报道的前列腺抗原(PA)检测方法,开发出一种反向酶联免疫吸附测定法(ELISA),用于检测循环中的PA结合球蛋白(PABG)。我们分析了取自正常男性、正常女性、非前列腺癌男性患者、良性前列腺增生患者以及不同阶段前列腺癌患者的血清样本中的PABG。结果显示,只有前列腺癌晚期患者的PABG水平升高。然后通过PA亲和层析从前列腺癌患者血清中分离出PABG。经免疫扩散和免疫电泳分析,该PABG制剂与纯化的PA、抗PA异种抗体及抗人IgG发生反应。采用免疫过氧化物酶技术,PABG在前列腺导管上皮细胞中呈阳性染色,而在所有其他检测组织中呈阴性染色。通过抗PA亲和层析还分离出了另一种PABG制剂,它与抗PA和抗人IgG发生反应,但不与纯化的PA反应。这些PABG制剂分别进行高效液相色谱进一步纯化。得到了分子量大于240K、150K和34K的三个主要蛋白峰。这些结果表明,前列腺癌转移患者体内存在与PA反应的循环IgG抗体,部分以与PA形成复合物的形式存在,并与前列腺导管上皮成分发生特异性反应。