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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。
Objective To establish a relatively simple method for measuring the affinity of erythrocyte antibody in order to study the effect of affinity on the detection of erythrocyte antibody. Methods According to the affinity formula, when the reaction between erythrocytes and antibody reached equilibrium, the binding constant (Ka) between the antibody and the corresponding erythrocyte, ie, affinity, was calculated by measuring the concentration of free antibody and calculating the ratio of antigen bound to unbound antibody on erythrocytes. Using quantitative R1R1 erythrocytes to absorb high-titer human anti-D, first determine the amount of saturated anti-D, and then measure the free antibody concentration after the different concentrations of anti-D were quantitated, and calculate the concentration of erythrocyte bound and unbound anti-D D antigen ratio derived human anti-D and R1R1 cell binding reaction constants. Results The affinity of human anti-D and R1R1 cells used in the assay was (2.1-7.6) × 108. The experiment showed that the lower the amount of red blood cell bound antibody, the higher the affinity. The affinity of one monoclonal IgG anti-D detected in parallel was about 4.6 × 109, which was significantly higher than that of human anti-D. Conclusion The established method for measuring the affinity of erythrocyte antibody is more convenient and reduces the possibility of deviation. It is suitable for the study of basic, clinical and testing methods.