脾内一次性免疫莸取抗人体肺癌单克隆抗体

MONOCLONAL ANTIBODY AGAINST HUMAN LUNG ADENOCARCINOMA BY INTRASPLENIC PRIMARY IMMUNIZATION

用人体肺腺癌细胞(LAX)脾内一次性免疫BALB/C小鼠后,将脾细胞与小鼠骨髓瘤细胞(NSO)融合,融合率为80％,对LAX细胞反应阳性率为20％。以酶联免疫吸附试验,免疫荧光试验和免疫组织化学试验,检测其与正常人体细胞,人体肿瘤细胞和人体肺腺癌的反应,发现获得的ALA-05单克隆抗体与正常人体细胞无交叉反应,与多株人体其他肿瘤细胞也无反应,而与人体肺腺癌则呈阳性反应,且为抗膜抗体。免疫扩散法证明其是IgG_(2a)。

BALB/C mice were immunized with human lung adenocarcinomacell LAX by intraspleen route. The splenocytes were fused with myeloma cellNSO. Hybridoma cells grew in 80% wells of 8 cell culture plates. Twentypercent of hybridoma clones reacted with LAX cells in the primary screening. ELISA and immunofluorescence stain showed that resulting monoclonal antibodyALA-05 (Immunodiffusion indicated IgG(2a)) could recognize membrane antigenon human lung cancer very strongly, but did not react with various normaltissues and several other kinds of human tumor cell lines.