Nelson D P, Griswold W R
J Clin Lab Immunol. 1986 Oct;21(2):101-6.
Antigen-antibody binding experiments were performed by choosing antigen excess starting conditions and then diluting both the 125I-BSA antigen and anti-BSA proportionately so that the ratio between the reactants remained constant. The fraction antigen bound was measured at each dilution. Binding data were analyzed by computer using non-linear least squares regression to determine the affinity and affinity distribution of different antisera. Early anti-BSA was found to have a unimodal distribution with a binding constant in the range of 10(6)M-1. Intermediate anti-BSA had a bimodal distribution: 1/3 high affinity (1.0 X 10(9] and 2/3 low affinity (3.4 X 10(6)M-1). Late and hyperimmune rabbit BSA antibodies had unimodal affinity distributions with binding constants varying between 1.7 and 2.9 X 10(10)M-1. Antibody affinity can be readily determined by computer analysis of binding curves obtained in constant antigen excess conditions.
抗原 - 抗体结合实验通过选择抗原过量的起始条件来进行，然后按比例稀释¹²⁵I - BSA抗原和抗BSA，使得反应物之间的比例保持恒定。在每次稀释时测量结合的抗原分数。使用非线性最小二乘法回归通过计算机分析结合数据，以确定不同抗血清的亲和力和亲和力分布。发现早期抗BSA具有单峰分布，结合常数在10⁶M⁻¹范围内。中期抗BSA具有双峰分布：1/3高亲和力（1.0×10⁹）和2/3低亲和力（3.4×10⁶M⁻¹）。晚期和超免疫兔BSA抗体具有单峰亲和力分布，结合常数在1.7至2.9×10¹⁰M⁻¹之间变化。通过在恒定抗原过量条件下获得的结合曲线的计算机分析，可以很容易地确定抗体亲和力。
