抗人肝癌单克隆抗体HL_2的制备及鉴定

Preparation and Identification of Anti-Human Hepatocellular Carcinoma Monoclonal Antibody HL_2

用肝癌细胞株QGY-7703、BEL-7402、SMMC-7721以贯序法免疫BALB/c小鼠,获得一株分泌肝癌单抗的杂交窟株HL_2,其染色体数目为97—105。单抗HL_2系IgG_2b亚类。吸收实验及阻断实验证明HL_2抗原与CEA、AFP、HBsAg、HBcAg及HBeAg无免疫同源性。ABC法和ELISA法说明单抗HL_2与四株肝癌细胞、六例肝癌组织均呈明确阳性反应,除与SGC-7901、H_(128)、K_(562)、Hela细胞株及部分胚胎肝脏,胚胎结肠有较弱性反应外,与肝癌旁组织、正常肝脏、其它肿瘤组织和细胞株、二种正常人细胞及其它胚胎组织无反应。显示了单抗HL_2的特异性较好、阳性率较高,是一个有应用前景的单抗。

The human hepatocellular carcinoma (HCC) cell lines, QGY-7703, BEL-7402, SMMC-7721 were used to immunize BALB/c mouse in a sequential procedure and one hybridoma, HL2, producing monoclonal antibody (McAb) to HCC cells were established. The amounts of chromosomes of HL2 cells are from 97 to 105. McAb HL2 were found to be of the IgG2b isotye using double diffusion.The result suggests that McAb HL2 identify a HCC-associated antigen, which may be named HL2 antigen. HL2 antigen was located mostly on the membrane and cytoplasm of HCC that showed by avidin-biotin peroxidose complex (ABC) technique. Blocking and absorbing experiments revealed the HL2 antigen had no immuno-homogenicity with those antigens such as HBsAg, HBeAg, HBcAg, AFP and CEA. Specificity of McAb HL2 was tested by enzyme-linked immunosorbent assay (ELISA) and ABC technique. McAb HL2 reacted definitely with 4 of 4 HCC lines and lightly with 4 of 12 other different human tumor lines, but not with the others and 2 normal human cells. McAb HL2 was also positive in 6 of 6 HCC patients but negative in 5 liver tissues nearby the HCC tissues, 3 normal adult liver tissues and 6 other human tumor tissues.Eleven kinds of fetal tissues were also found to be negative to McAb HL2,except for some reactions to 3 of 5 the fetal liver tissues and 2 of 5 the fetal colon tissues. Our result suggests that McAb HL2 may be used in a bright prospect.