[抗血红蛋白δ珠蛋白链和β珠蛋白链高亲和力单克隆抗体的制备]
[Preparation of high-affinity monclonal antibody against hemoglobin delta globin chain and beta globin chain].
作者信息
Zhu Ping, Xiao Hai-yan, Chen Yi-guo, Xu Mei, Fu Ning
机构信息
Department of Immunology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Jun;26(6):556-9.
AIM
To prepare monclonal antibody (mAb) against both hemoglobin A2 (HbA2) and hemglobin A (HbA), this antiboy can bind to delta globin chain and beta globin chain, but not to gamma globin chain.
METHODS
Mice was immunized with recombinant Hb delta fusion protein, and hybridoma cells were generated by cell fusion techniques followed by screening with natural HbA2 and HbA separated by ion exchange chromatography. The purified monclonal antibody was identified by indirect ELISA, Western blot based on native PAGE and SDS-PAGE, surface plasmon resonance (SPR), flow cytometry and immunohistochemistry.
RESULTS
The monoclonal antibody against both HbA2 and HbA was obtained and designated as 2C9 that shows no binding to fetal hemoglobin (HbF), alpha globin chain and recombinant zetaglobin chain.
CONCLUSION
The mAb 2C9 was defined as specificity to hemoglobin delta globin chain and beta globin chain, which suggests that mAb 2C9 recognizes a common epitope on bothdeltaglobin chain and beta globin chain. This antibody would be expected to be an effective tool in research and clinical practice in hemoglobinopathies.
目的
制备针对血红蛋白A2（HbA2）和血红蛋白A（HbA）的单克隆抗体（mAb），该抗体可与δ珠蛋白链和β珠蛋白链结合，但不与γ珠蛋白链结合。
方法
用重组Hbδ融合蛋白免疫小鼠，通过细胞融合技术产生杂交瘤细胞，然后用离子交换色谱法分离的天然HbA2和HbA进行筛选。通过间接ELISA、基于天然PAGE和SDS-PAGE的Western印迹、表面等离子体共振（SPR）、流式细胞术和免疫组织化学对纯化的单克隆抗体进行鉴定。
结果
获得了针对HbA2和HbA的单克隆抗体，命名为2C9，其与胎儿血红蛋白（HbF）、α珠蛋白链和重组ζ珠蛋白链无结合。
结论
单克隆抗体2C9被定义为对血红蛋白δ珠蛋白链和β珠蛋白链具有特异性，这表明单克隆抗体2C9识别δ珠蛋白链和β珠蛋白链上的共同表位。该抗体有望成为血红蛋白病研究和临床实践中的有效工具。
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