抗人ζ珠蛋白链不同表位单克隆抗体的制备和鉴定

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (11): 1394-1397.

抗人ζ珠蛋白链不同表位单克隆抗体的制备和鉴定

唐磊¹, 朱平¹, 罗琛², 徐湘民², 富宁¹

1. 南方医科大学免疫教研室, 广东, 广州, 510515;
2. 南方医科大学医学遗传学教研室, 广东, 广州, 510515

出版日期:2005-11-20 发布日期:2005-11-20
基金资助:
收稿日期:2005-4-14。
基金项目:广东省科技计划项目(2004-A30801003)
作者简介:唐磊(1976-),男,南方医科大学在读硕士研究生
通讯作者:富宁,电话:020-61648221

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

TANG Lei¹, ZHU Ping¹, LUO Chen², XU Xiang-ming², FU Ning¹

1. 南方医科大学免疫教研室, 广东, 广州, 510515;
2. 南方医科大学医学遗传学教研室, 广东, 广州, 510515

Online:2005-11-20 Published:2005-11-20

摘要/Abstract

摘要： 目的制备和鉴定抗ζ珠蛋白链单克隆抗体。方法用纯化的重组ζ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ζ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1A12、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ζ珠蛋白链,且其中1A12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。

Abstract: Objective To prepare and identify the monoclonal antibodies (mAb) against human ζ globin chain. Methods BALB/c mice were immunized with purified recombinant ζ 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 ζ 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-ζ serum. Results and Conclusion Three hybridoma cell lines secreting anti-ζ 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 ζ and natural ζ globin from hemolysate of --^SEA gene carrier. In addition, 1A12 and 3H9 mAbs could recognize different epitopes on ζ globin, suggesting the possibility of developing a detection system for screening alpha-thalassemia with these mAbs.

中图分类号:

R392.11

唐磊¹, 朱平¹, 罗琛², 徐湘民², 富宁¹. 抗人ζ珠蛋白链不同表位单克隆抗体的制备和鉴定[J]. 南方医科大学学报, 2005, 25(11): 1394-1397.

TANG Lei¹, ZHU Ping¹, LUO Chen², XU Xiang-ming², FU Ning¹. Preparation and identification of monoclonal antibodies against different epitopes on human ζ globin chain[J]. Journal of Southern Medical University, 2005, 25(11): 1394-1397.

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