Backen A C, Cummings J, Mitchell C, Jayson G, Ward T H, Dive C
CR-UK Translational Angiogenesis Group, Paterson Institute for Cancer Research, University of Manchester, Manchester, UK.
J Immunol Methods. 2009 Mar 15;342(1-2):106-14. doi: 10.1016/j.jim.2009.01.003. Epub 2009 Jan 25.
Validated assays of circulating biomarkers of angiogenesis to predict and determine the efficacy of vascular-targeted anticancer drugs would facilitate successful drug development. Multiple biomarker candidates exist and a multiplex approach was sought to minimise the requisite patient blood volume and to aid selection of those biomarkers with greatest potential clinical utility. Validation of the SearchLight multiplex ELISA platform comprising two multiplex assays of nine potential angiogenesis biomarkers was conducted (plex 1; VEGF R1 and R2, IL-8, KGF, PlGF; plex 2; PDGFbb, HGF, FGFb and VEGF). The study focused on instrument qualification, analyte specificity within the multiplex format, assay precision and reproducibility. No evidence was found within the multiplex that signals output from one analyte impinged on another or that antibody cross-reactivity occurred. Spike recovery for 5 between-experiment repeats was within +/-15% of input values for 7 of the 9 multiplexed analytes, with a coefficient of variation (CV) of <20% for 6 of the 9 analytes. Plasma samples from 8 ovarian cancer patients (who were not receiving therapy) were assessed using the two multiplexes on this platform to explore the likely baseline variability in this disease context. This study suggests that the platform and the multiplex approach will be useful to evaluate pharmacodynamic responses to vascular targeted therapy in early clinical trials.
验证循环血管生成生物标志物的检测方法以预测和确定血管靶向抗癌药物的疗效，将有助于成功开展药物研发。目前存在多种生物标志物候选物，因此寻求一种多重检测方法，以尽量减少所需的患者血容量，并帮助选择具有最大潜在临床应用价值的生物标志物。对SearchLight多重ELISA平台进行了验证，该平台包括对9种潜在血管生成生物标志物的两种多重检测（检测组1；血管内皮生长因子受体1和2、白细胞介素-8、角质形成细胞生长因子、胎盘生长因子；检测组2；血小板衍生生长因子bb、肝细胞生长因子、碱性成纤维细胞生长因子和血管内皮生长因子）。该研究重点关注仪器鉴定、多重检测形式内分析物的特异性、检测精度和重现性。在多重检测中未发现证据表明一种分析物输出的信号会影响另一种分析物，也未发现抗体交叉反应。9种多重分析物中有7种在5次实验重复间的加标回收率在输入值的±15%以内，9种分析物中有6种的变异系数（CV）<20%。使用该平台上的两种多重检测方法对8名未接受治疗的卵巢癌患者的血浆样本进行了评估，以探索在这种疾病背景下可能的基线变异性。这项研究表明，该平台和多重检测方法将有助于在早期临床试验中评估对血管靶向治疗的药效学反应。
