抗人Ⅳ型胶原单抗研制及其特性

The Preparation and Characterization of Two Monoclonal Antibodies to Human Collagen Type IV

本文用胃蛋白酶提取的人胎盘Ⅳ型胶原免疫Balb/c小鼠,免疫脾脏细胞与小鼠骨髓瘤细胞融合后,经有限稀释法克隆获得两株分泌抗人Ⅳ型胶原单抗的杂交瘤细胞株CO_5和CO_6。用小鼠腹水和杂交瘤细胞培养上清纯化制备的两种单抗均属小鼠IgGl,K键,分别针对Ⅳ型胶原分子上的两个不同抗原决定簇。两种单抗特异性强,与人Ⅰ型、Ⅱ型、Ⅲ型、Ⅴ型、Ⅵ型、Ⅷ型胶原、Ⅳ型胶原7S区及人白蛋白等相关蛋白无明显交叉反应,且单抗效价高,亲和力强。

Balb/c mouse was immunized with pepsin-extracted collagen type Ⅳ from human placenta. The spleen cells of the mouse were fused with NS-1 mouse myoloma cells. Two monoclonal antibody secreting hybridoma cell lines, CO5 and CO6 ,were obtained through the limiting dilution. Both CO5 and CO6 McAbs from mouse ascites or hybridoma cell culture supernatants were iso-typed to mouse IgG1 , K Chain and directed against two different determinants on human collagen type Ⅳ molecule. No crossreactivities were found with other human collagens of type Ⅰ , Ⅱ ,Ⅲ , Ⅴ , Ⅵ , Ⅶ, 7s domains of collagen type Ⅳ and human albumin. Both monoclonal antibodies have high titers and affinities to the antigen.