Becker C, Wigheden I, Lilja H
Department of Clinical Chemistry, Lund University, University Hospital, Malmö, Sweden.
Tumour Biol. 1999;20 Suppl 1:13-7. doi: 10.1159/000056524.
Prostate-specific antigen (PSA) is the most widely used marker of prostate cancer. Assays for PSA are based on anti-PSA antibodies, and the characterization and selection of these antibodies is important for determining their optimum performance. In our study, we characterized the reactivity of 53 antibodies, submitted to the ISOBM TD-3 PSA Workshop, using free PSA, PSA complexed to alpha1-antichymotrypsin (ACT) and purified ACT. Immunoblotting was performed after native agarose gel or reducing sodium dodecyl polyacrylamide gel electrophoresis. Immunoblotting after agarose gel electrophoresis revealed 10 antibodies that recognized only the free form of PSA, and 43 antibodies that detected both free PSA and PSA-ACT. Immunoblotting of reducing sodium dodecyl-polyacrylamide gels showed the linear or conformation-dependent nature of the epitopes. Two antibodies specific for free PSA and 18 antibodies that recognized both free PSA and PSA-ACT complex recognized linear epitopes. Moreover, 7 antibodies also detected fragmented forms of PSA.
前列腺特异性抗原（PSA）是前列腺癌最广泛使用的标志物。PSA检测基于抗PSA抗体，这些抗体的特性鉴定和选择对于确定其最佳性能很重要。在我们的研究中，我们使用游离PSA、与α1-抗糜蛋白酶（ACT）复合的PSA以及纯化的ACT，对提交给国际生物大分子标准品委员会（ISOBM）TD-3 PSA研讨会的53种抗体的反应性进行了表征。在天然琼脂糖凝胶或还原十二烷基硫酸钠聚丙烯酰胺凝胶电泳后进行免疫印迹分析。琼脂糖凝胶电泳后的免疫印迹分析显示，有10种抗体仅识别游离形式的PSA，43种抗体可检测游离PSA和PSA-ACT。还原十二烷基硫酸钠-聚丙烯酰胺凝胶的免疫印迹分析显示了表位的线性或构象依赖性性质。两种针对游离PSA的特异性抗体和18种识别游离PSA和PSA-ACT复合物的抗体识别线性表位。此外，7种抗体还检测到了PSA的片段形式。
