Gallee M P, van Vroonhoven C C, van der Korput H A, van der Kwast T H, ten Kate F J, Romijn J C, Trapman J
Prostate. 1986;9(1):33-45. doi: 10.1002/pros.2990090107.
For production of monoclonal antibodies (McAbs), hybrid cells were prepared by fusion of spleen cells of BALB/c mice immunized with the human prostatic cancer cell line PC-82 and the P3-X6(3)Ag8.653 murine myeloma cell line. Supernatants of approximately 500 hybrid clones were screened for prostate specific antibodies using an ELISA on PC-82 cells. A selection of antibodies was further tested for their specificity on a large series of different tissues. A broad cross reactivity pattern was obtained. Most cross reactivity was with pancreatic tissue, kidney, and bowel. One antibody turned out to react with prostate stromal cells. Two McAbs (ER-Pr 1 and ER-Pr 2) reacted solely with prostatic epithelium. Monoclonal antibody affinity chromatography combined with SDS-PAGE showed that both antibodies were directed against a 35-kD protein. Immunoblotting revealed that this protein is identical to prostatic antigen (PA). The epitope detected by ER-Pr 1 and ER-Pr 2 was largely preserved after formalin-fixation of prostatic tissues which renders these antibodies very suitable for routine examination of tissue sections.
为制备单克隆抗体（McAbs），将用人前列腺癌细胞系PC - 82免疫的BALB/c小鼠的脾细胞与P3 - X6(3)Ag8.653小鼠骨髓瘤细胞系融合，制备杂交细胞。使用针对PC - 82细胞的酶联免疫吸附测定（ELISA），对约500个杂交克隆的上清液进行前列腺特异性抗体筛选。进一步对一系列抗体在大量不同组织上的特异性进行检测。获得了广泛的交叉反应模式。大多数交叉反应发生在胰腺组织、肾脏和肠道。有一种抗体被证明与前列腺基质细胞发生反应。两种单克隆抗体（ER - Pr 1和ER - Pr 2）仅与前列腺上皮发生反应。单克隆抗体亲和层析结合十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）显示，这两种抗体均针对一种35 kDa的蛋白质。免疫印迹表明，该蛋白质与前列腺抗原（PA）相同。前列腺组织经福尔马林固定后，ER - Pr 1和ER - Pr 2检测到的表位基本保留，这使得这些抗体非常适合用于组织切片的常规检查。
