Nonaka N, Hirai H
Tumour Laboratory, Tokyo.
Jpn J Cancer Res. 1994 Apr;85(4):372-7. doi: 10.1111/j.1349-7006.1994.tb02369.x.
A component exhibiting toxicity to B cell hybridoma cells was isolated and purified from fetal calf serum (FCS) by immunoaffinity chromatography using a monoclonal antibody (mAb) which reacted with the high-molecular-weight glycoprotein (6B3.Ag) recognized by a mAb, 6B3, to human large cell lung carcinoma cells (HLC-2). The component (FCS-6B3.Ag) was a high-molecular-weight antigen (approximately 1,000,000), consisting mainly of 76,000 subunits. FCS-6B3.Ag showed the same mobility in the pre-beta globulin region as that of 6B3.Ag on electrophoresis in 1.2% agarose gel. When FCS-6B3.Ag was analyzed by double immunodiffusion, it reacted with anti-6B3.Ag antiserum and the precipitin line fused partially with that formed between 6B3.Ag and anti-6B3.Ag antiserum. FCS-6B3.Ag was found to be toxic to hybridoma cells (anti-6B3.Ag, anti-alpha-fetoprotein, anti-carcinoembryonic antigen or anti-C-reactive protein mAb producing cells) specifically in vitro at 5 micrograms/ml. The antigen also strongly suppressed their growth. The toxic effect of FCS-6B3.Ag appeared immediately after addition, and death of the target cells was complete only after 36-48 h. However, the antigen exhibited only weak suppression of Ig-non-secretory mouse myeloma (P3U1), thymic lymphoma (EL4) of mastocytoma (P815) cell growth. Five lots of FCS contained 2.1 to 4.1 micrograms/ml of FCS-6B3.Ag.
使用与抗人大细胞肺癌细胞（HLC - 2）的单克隆抗体6B3识别的高分子量糖蛋白（6B3.Ag）反应的单克隆抗体，通过免疫亲和色谱从胎牛血清（FCS）中分离并纯化出一种对B细胞杂交瘤细胞具有毒性的成分。该成分（FCS - 6B3.Ag）是一种高分子量抗原（约1,000,000），主要由76,000亚基组成。在1.2%琼脂糖凝胶电泳中，FCS - 6B3.Ag在前β球蛋白区域的迁移率与6B3.Ag相同。当通过双向免疫扩散分析FCS - 6B3.Ag时，它与抗6B3.Ag抗血清反应，沉淀线与6B3.Ag和抗6B3.Ag抗血清之间形成的沉淀线部分融合。发现FCS - 6B3.Ag在体外对杂交瘤细胞（产生抗6B3.Ag、抗甲胎蛋白、抗癌胚抗原或抗C反应蛋白单克隆抗体的细胞）具有特异性毒性，浓度为5微克/毫升。该抗原也强烈抑制它们的生长。FCS - 6B3.Ag添加后立即出现毒性作用，靶细胞仅在36 - 48小时后完全死亡。然而，该抗原对Ig非分泌性小鼠骨髓瘤（P3U1）、胸腺淋巴瘤（EL4）或肥大细胞瘤（P815）细胞生长仅表现出微弱的抑制作用。五批FCS中FCS - 6B3.Ag的含量为2.1至4.1微克/毫升。
