CAJ | 학술논문

目的研制血管内皮生长因子(vascular endothelial growth factor,VEGF)检测试剂盒(酶标法).方法以一株特异性抗VEGF单克隆抗体作为捕获抗体,另一株单克隆抗体作为检测抗体,通过棋盘滴定法确定两抗体的最佳浓度组合.制备冻干VEGF165参考品,并对样本稀释液、封闭液等实验条件进行优化.检测自制试剂盒的特异性并对其进行初步应用.结果棋盘滴定法确定捕获抗体的质量浓度为10μg/ml,检测抗体的稀释倍数为1:20 000.冻干VEGF165的质量浓度为720～880 pg/ml,样本间变异系数<10%.以5%牛血清白蛋白作为封闭液和样本稀释液最为合适.自制试剂盒与10种细胞因子均无交叉反应,具有较高的特异性和灵敏度.结论成功研制出VEGF检测试剂盒.
목적 연제혈관내피생장인자(vascular endothelial growth factor,VEGF)검측시제합(매표법).방법 이일주특이성항VEGF단극륭항체작위포획항체,령일주단극륭항체작위검측항체,통과기반적정법학정량항체적최가농도조합.제비동간VEGF165삼고품,병대양본희석액、봉폐액등실험조건진행우화.검측자제시제합적특이성병대기진행초보응용.결과 기반적정법학정포획항체적질량농도위10μg/ml,검측항체적희석배수위1:20 000.동간VEGF165적질량농도위720～880 pg/ml,양본간변이계수<10%.이5%우혈청백단백작위봉폐액화양본희석액최위합괄.자제시제합여10충세포인자균무교차반응,구유교고적특이성화령민도.결론 성공연제출VEGF검측시제합.
Objective To develop a vascular endothelial growth factor (VECF) assay kit. Methods Two anti-VEGF monoclonal antibodies were used as capture and detection antibodies, respectively, and optimum concentration combination of the antibodies was determined by chessboard titration. Freeze-dried VEGF165 reference material was prepared. Sample diluent and blocking solution were screened. The kit was tested for specificity and applied preliminarily. Results The concentration and dilution multiple of the capture antibody and detection antibody were identified as 10 μg/ml and 1:20 000, respectively. The concentration of freeze-dried VEGF165 reference material was 720- 880 pg/ml and the variation coefficient was < 10%. 5% bovine serum albumin was chosen both as sample diluent and as blocking solution. No cross-reactions between the kit and 10 other cytokines were found. The kit had high sensitivity and specificity. Conclusion The VECF assay kit was successfully developed.