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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。
Objective To obtain specific aptamers with the ability to antagonize RhD antibodies. Methods SELEX technology was used to select the suitable aptamers which can bind to RhD antibody from the 82 nt random single-stranded DNA library synthesized in vitro. The affinity and specificity of nucleic acid aptamers binding to RhD antibodies were detected by fluorescent labeling, and the dose-response relationship of neutralizing RhD antibodies was analyzed. Results The dissociation constants of 4 aptamers reached nmol / L level. Among them, aptamer 3 showed the strongest specificity, followed by No.1, while Nos. 2 and 4 showed no specificity. 1 and 3 nucleic acid aptamers used in combination, the concentration of 50 pmol / L, can completely neutralize RhD antibodies. Conclusion The nucleic acid aptamers that specifically bind to RhD antibodies were successfully obtained by using random single-stranded oligonucleotide libraries. The obtained aptamers have the ability of antagonizing RhD antibodies.