芍药苷单克隆抗体的特异性检测

Immunospecificity Detections of Lab-prepared Anti-paeoniflorin Monoclonal Antibody

对已制备的中药活性成分芍药苷的单克隆抗体的效价、敏感度、稳定性以及其与若干化学结构类似物的交叉反应性等进行了测定,据此评价单克隆抗体的免疫反应特异性,为药用植物活性成分的单克隆抗体的制备及其应用研究提供现有条件下的试验方法和依据.检测结果显示免疫小鼠腹水的单抗效价达到1∶256 000,来自细胞株P1E6和P5h10的单抗的灵敏度分别为0.080、0.124μg/mL,亲和常数1.819×10-7、2.815×10-7mol/L.同时,所选5种化学结构类似物均显示极低的交叉反应性,证明了芍药苷单克隆抗体的高度特异性.

Immunospcificity detections of the lab-prepared anti-paeoniflorin monoclonal antibody （anti-PF MAb） were carried out to establish an enzyme-linked immunosorbent assay （ELISA） for bio-active compounds ,paeoniflorin, from traditional Chinese medicines. The results showed that the titer ofasckes containing anti-PF MAbs from both cell strands P1E6 and P5H10 reached 1 ： 256 000. The sensitivity was 0.080 and0. 124 μg/mL, respectively. The affinity constant was 1. 819 × 10^-7 mol/L for MAb of P1E6 and 2. 815 × 10^-7 mol/L of P5H10, respectively. Five chemically-structural analogues showed very low cross-reactivity with PF, suggesting a very high immunospecificity of the prepared anti-PF MAbs and the feasibility in determination of PF from Chinese materia medica.