人源抗-D亲和力测定方法的建立及其验证

发布时间：2018-11-08 16:07

【摘要】：目的建立1种相对简便的测量红细胞抗体亲和力方法,以之研究亲和力因素对红细胞抗体检测的影响。方法根据亲和力公式,在红细胞与抗体反应达到平衡时,通过测量游离抗体浓度以及计算红细胞表面结合与未结合抗体的抗原比例,可计算出抗体与相应红细胞的结合常数(Ka)即亲和力。应用定量的R1R1红细胞吸收高效价人源抗-D,先测定饱和吸收抗-D的量,再测量不同浓度抗-D被定量红细胞吸收后游离抗体浓度,计算红细胞结合与未结合抗-D的D抗原的比值,推导出人源抗-D与R1R1细胞反应的结合常数。结果检测中使用的人源抗-D与R1R1细胞反应亲和力为(2.1-7.6)×108;实验显示,红细胞结合抗体量越少,亲和力相对越高。平行检测的1例单克隆Ig G抗-D的亲和力约为4.6×109,明显高于人源抗-D。结论所建立的测量红细胞抗体亲和力方法更为简便,减少了出现偏差的机会,适用于基础、临床以及检测的方法研究。
[Abstract]:Objective to establish a relatively simple method for the determination of erythrocyte antibody affinity and to study the effect of affinity factors on erythrocyte antibody detection. Methods according to the formula of affinity, when the reaction between erythrocyte and antibody reached equilibrium, the concentration of free antibody and the antigen ratio of surface binding and unbinding antibody of erythrocyte were calculated by measuring the concentration of free antibody. The binding constant (Ka) of the antibody and the corresponding red blood cell can be calculated. Using the quantitative R1R1 red blood cell absorption high titer human anti-D, the saturated absorption anti-D was measured first, then the free antibody concentration after the different concentration of anti-D was absorbed by the quantitative red blood cell was measured. The ratio of red blood cell binding to unbound D antigen was calculated and the binding constant of human anti D to R1R1 cells was derived. Results the affinity of human anti-D to R1R1 cells was (2.1-7.6) 脳 10 ~ (8), which showed that the lower the amount of erythrocyte binding antibody was, the higher the affinity was. The affinity of monoclonal Ig G anti-D was 4.6 脳 109, which was significantly higher than that of human anti-D. Conclusion the established method for measuring the affinity of red blood cell antibodies is more convenient and less prone to deviation. It is suitable for basic, clinical and detection methods.
【作者单位】：上海市血液中心;
【基金】：上海市公共卫生重点学科建设项目(15GWZK0501)
【分类号】：R457.1

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