抗人Ⅲ型胶原单克隆抗体的制备与正常组织定位

PREPARATION OF MONOCLONAL ANTIBODIES TO HUMAN TYPE Ⅲ COLLAGEN AND THEIR LOCALIZATION IN NORMAL TISSUES

用新鲜人肝癌组织细胞悬液免疫Balb/c小鼠,取小鼠脾细胞与小鼠骨髓瘤细胞系Sp2/0融合,获得两株抗人胶原蛋白单克隆抗体CAb-15和CAb-16。应用Western blotting方法对两株抗体的抗原决定簇进行了鉴定。结果表明,CAb-15的抗原决定簇位于α_1[Ⅲ]链,CAb-16位于Ⅲ型胶原的三螺旋结构区。应用CAb-15和CAb-16对在福尔马林中固定和石蜡包埋的正常肝、肾、主动脉、皮肤、软骨和胎盘等组织进行了免疫组织化学定位。结果表明两株Ⅲ型胶原抗体均可用于免疫组织化学研究。

Monoclonal antibodies, CAb-15 and CAb-16, were produced by using of spleen cells from Balb/c mice immunized with cell suspension of fresh hepatoma tissues with mouse myeloma Sp 2/0 cells. Western blotting showed that the antigen determinant of CAb-15 was situated on the α_1 [Ⅲ] polypeptide chains and that of CAb-16 on the triple helical domain. The monoclonal antibodies were also used for immunohistochemical localization by ABC method in formalin-fixed and paraffin-embedded sections of normal tissues from liver, kidney, aorta, skin, cartilage, placenta, stomach, breast and granulation tissues. The results showed that these antibodies are useful in immunohistochemical study for collagen.