抗人ξ珠蛋白链不同表位单克隆抗体的制备和鉴定

目的制备和鉴定抗ξ珠蛋白链单克隆抗体。方法用纯化的重组ξ珠蛋白链免疫Balb／c小鼠，取其脾细胞与NS-1细胞融合制备杂交瘤，经重组的ξ珠蛋白链筛选和3次克隆化，从制备的腹水中纯化单克隆抗体。结果和结论获得3株杂交瘤细胞IA12、3H9及4D11，其分泌单抗的亚型分别为IgG2a、IgG1和IgG1；双抗体夹心ELISA结果证明3株单抗均可结合重组的及东南亚（--^SEA）缺失型地中海贫血基因携带者血中的ξ珠蛋白链，且其中IA12和3H9识别的位点为不同表位。这三株抗体将为筛选地中海贫血基因携带者的免疫检测提供有力工具。

Preparation and identification of monoclonal antibodies against different epitopes on human zeta globin chain]

OBJECTIVE: To prepare and identify the monoclonal antibodies (mAb) against human zeta globin chain. METHODS: BALB/c mice were immunized with purified recombinant zeta globin chain, and the hybridomas were generated by fusion of mouse spleen cells and myeloma cells NS-1. After three fusions and successive cloning, 3 hybridoma cell lines secreting the mAb against zeta were obtained and the antibodies were purified from the ascites, followed by identification with indirect enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA in combination with rabbit anti-zeta serum. RESULTS AND CONCLUSION: Three hybridoma cell lines secreting anti-zeta mAb were established, designated as 1A12, 3H9 and 4D11, respectively. Both of 3H9 and 4D11 mAbs belonged to IgG1 isotype and mAb 1A12 to IgG2a. All the mAbs could bind specifically to recombinant zeta and natural zeta globin from hemolysate of --(SEA) gene carrier. In addition, 1A12 and 3H9 mAbs could recognize different epitopes on zeta globin, suggesting the possibility of developing a detection system for screening alpha-thalassemia with these mAbs.