Wong J H, Xu S H, Saxton R E, Gupta R K, Morton D L
Division of Surgical Oncology, UCLA School of Medicine 90024.
J Surg Res. 1990 Jun;48(6):539-46. doi: 10.1016/0022-4804(90)90227-s.
A murine monoclonal antibody of the subclass IgG2b, designated MAb JSI, was produced utilizing standard hybridoma technology. Female BALB/c mice were immunized with a fetal antigen that had been partially purified from the spent serum-free culture media of a human melanoma cell line. Utilizing MAb JSI, the antigen was isolated from serum of melanoma patients by affinity chromatography utilizing an acid elution and studied following exposure to trypsin, protease, and heat in an enzyme-linked immunosorbent assay (ELISA). The antigenic activity was destroyed following treatment with trypsin and protease as well as by exposure to heat (100 degrees C). By immunoperoxidase staining, MAb JSI reacted with melanoma, carcinoma, and sarcoma cell lines, but not with cell lines derived from normal skin or lungs. Affinity-isolated antigen was subjected to polyacrylamide gel electrophoresis and stained with Coomassie blue. Under dissociating conditions, a band in the 31,000 to 42,700 Da range was identified that was shown to be reactive with MAb JSI in ELISA. The antigenic determinant defined by MAb JSI appears to be a protein, with expression on a number of malignant tissues.
利用标准杂交瘤技术制备了一种亚类为IgG2b的鼠单克隆抗体，命名为单克隆抗体JSI。用从人黑色素瘤细胞系的无血清培养液中部分纯化的胎儿抗原来免疫雌性BALB/c小鼠。利用单克隆抗体JSI，通过酸洗脱亲和层析从黑色素瘤患者血清中分离出该抗原，并在酶联免疫吸附测定（ELISA）中对其进行胰蛋白酶、蛋白酶和加热处理后进行研究。经胰蛋白酶和蛋白酶处理以及加热（100℃）后，抗原活性被破坏。通过免疫过氧化物酶染色，单克隆抗体JSI与黑色素瘤、癌和肉瘤细胞系发生反应，但与源自正常皮肤或肺的细胞系不发生反应。对亲和分离的抗原进行聚丙烯酰胺凝胶电泳并用考马斯亮蓝染色。在变性条件下，鉴定出一条分子量在31,000至42,700道尔顿范围内的条带，该条带在ELISA中显示与单克隆抗体JSI有反应性。单克隆抗体JSI所定义的抗原决定簇似乎是一种蛋白质，在多种恶性组织中表达。
