ELISA法间接检测血管内皮生长因子抑制剂的相对结合力

目的采用ELISA法间接检测血管内皮生长因子抑制剂(VEGF Trap)的相对结合力。方法以固定量的VEGF165与VEGF Trap结合,通过双抗体夹心ELISA法检测未结合的、游离的VEGF165,进而求得VEGFTrap的半数抑制浓度(IC50),计算其与反应体系中参比品的IC50之比,即为VEGFTrap的相对结合力。结果同一批次的3支VEGF Trap样品分别经3次测定,相对结合力分别为(100.86±9.43)%、(88.46±13.94)%和(82.66±9.60)%,均值为(90.66±12.60)%,符合该制品质量标准的要求。3次试验的变异系数分别为9.35%、15.76%和11.61%。结论 ELISA法精密性良好,结果客观,影响因素少,可用于VEGF Trap相对亲和力的常规检测。

Determination of Relative Binding Affinity of Vascular Endothelial Growth Factor Trap by Indirect ELISA

Objective To determine the relative binding affinity of vascular endothelial growth factor(VEGF)Trap by indirect ELISA.Methods The VEGF165 at a fixed amount was bound to VEGF Trap,and the free VEGF165 was determined by double antibody sandwich ELISA,based on which the median inhibition concentration(IC50)of VEGF Trap as well as its ratio to the IC50 of reference in reaction system,i.e.the relative binding affinity,were calculated.Results Each of the three VEGF Trap samples of the same batch was determined for 3 times,and the relative binding affinities were(100.86 ± 9.43)%,(88.46 ± 13.94)% and(82.66 ± 9.60)% respectively,at a mean of(90.66 ± 12.60)%,which met the requirement for quality control.The variation coefficients of three tests were 9.35%,15.76% and 11.61% respectively.Conclusion ELISA showed high precision,and its determination result was objective with few influencing factors,suggesting that the method might be used for routine determination of relative binding affinity of VEGF Trap.