非竞争ELISA法测定抗DR5单克隆抗体功能性亲和常数

目的:测定抗DR5单克隆抗体功能性亲和常数。方法:采用非竞争性ELISA固相法,经确定最佳抗原包板浓度、最佳抗原包板时间及最佳抗原与抗体结合反应时间后,得到DR5与抗DR5的两株单克隆抗体的抗原抗体结合反应曲线,计算抗DR5两株单抗的亲和常数。结果:抗DR5(366EC)的功能性亲和常数在2.63×109M-1水平,抗DR5(mDRA-6)的功能性亲和常数在1.004×109。结论:抗DR5与DR5结合能力较强,为今后进行免疫学检测和开发进行肿瘤靶向治疗提供了理论基础。

Measurement of function affinity of anti-DR5 McAb using non-competitive ELISA method

O b jective: To assess function affin ity of anti-DR5 M cAb in order to provide theoretical basis for the immunotherapy of cancer.M ethod s: W ith cogn izance of the advantages of solid phase m ethod in tim e consumption and conven ience,the functional affin ity of anti-DR5 M cAb was assessed w ith non-competitive ELISA m ethod.A fter a series of trials the best concentration and the best tim e of DR5 to cover the p late and the proper b ind ing tim e of DR5 to reach an equ ilibrium were affirm ed.W e p lotted the OD standard curve of the b ind ing reaction of DR5 using four grades of concentration ofDR5.W e got the consistency of anti-DR5 atOD50% through the standard curve,and then calcμlated the affin ity by Law ofM ass Action.R esμlts: The affin ity of anti-DR5 is 109M-1.C onc lu sion: The anti-DR5 wh ich we constructed in our Lab,can strongly b ind to the target antigen,thus provid ing a theoretical basis for its Immunological detection and c lin ical use.