RhD抗体ssDNA适配体的筛选与鉴定

发布时间：2018-01-22 16:46

本文关键词： 单链DNA 核酸适配体 RhD抗体指数富集的配体系统进化技术　出处：《中国输血杂志》2016年06期 　论文类型：期刊论文

【摘要】：目的获得具有拮抗RhD抗体能力的特异性核酸适配体。方法利用SELEX技术,从体外合成的82 nt随机单链DNA文库中,筛选出与RhD抗体特异结合的核酸适配体。采用荧光标记法检测核酸适配体与RhD抗体结合的亲和力及特异性,并对中和RhD抗体的剂量效应关系进行分析。结果通过筛选得到的4个核酸适配体解离常数达到nmol/L水平,其中3号核酸适配体特异性最强,其次为1号,而2号与4号无特异性。1号与3号核酸适配体联合使用,浓度分别为50 pmol/L时,可完全中和RhD抗体。结论利用随机单链寡核苷酸文库成功获得与RhD抗体特异性结合的核酸适配体,所获得的核酸适配体具有拮抗RhD抗体的能力。
[Abstract]:Objective to obtain a specific nucleic acid aptamer with the ability of antagonizing RhD antibody. Methods 82 NT random single-stranded DNA library was synthesized in vitro using SELEX technique. Nucleic acid aptamers that specifically bind to RhD antibody were screened. The affinity and specificity of the aptamer to RhD antibody were detected by fluorescence labeling method. The dose-effect relationship of neutralizing RhD antibody was analyzed. Results the dissociation constants of 4 nucleic acid aptamers obtained by screening reached the level of nmol/L, and the specificity of nucleic acid aptamer No. 3 was the strongest. The second was 1, but the 2 and 4 were not specific. When 1 and 3 aptamer were used together, the concentration was 50 pmol/L. Conclusion the nucleic acid aptamers specifically bound to RhD antibody were successfully obtained by using the random single-strand oligonucleotide library. The obtained aptamer has the ability to antagonize RhD antibody.
【作者单位】：深圳市血液中心;陕西省血液中心;广州中医药大学第二附属医院;
【基金】：深圳市科技计划项目(JCYJ20140403092619633)
【分类号】：R457.1
【正文快照】： proven to have the ability to neutralize Rh D antibody.Rh血型系统抗原可分为2类:由RHD基因编码的Rh D抗原与由RHCE基因编码的Rh Cc Ee抗原[1]。Rh D抗原的免疫性较强,是影响临床用血安全的重要因素,《临床输血技术规范》明确规定常规输血治疗前须检测ABO血型系统及Rh D抗

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