Wu Jie, Yan Feng, Tang Jinhai, Zhai Chun, Ju Huangxian
Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, Peoples Republic of China.
Clin Chem. 2007 Aug;53(8):1495-502. doi: 10.1373/clinchem.2007.086975. Epub 2007 Jun 28.
Automated and convenient multianalyte detection with high throughput is increasingly needed in clinical diagnosis. We developed a disposable 4-by-2 array for programmed simultaneous amperometric immunoassay of 4 tumor markers.
We used a screen-printed technique, 1-step immobilization method, and flow injection technique. We immobilized carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 as model analytes in a redox mediator-grafted, biopolymer-modified, screen-printed carbon electrode array to capture corresponding horseradish peroxidase-labeled antibodies in competitive immunoreactions. The simultaneous multianalyte immunoassay was automatically carried out to amperometrically monitor the mediator-catalyzed enzymatic response to hydrogen peroxide, which decreased in proportion to the concentrations of analytes in samples.
The multianalyte immunosensor array had a throughput of 60 samples/h and allowed simultaneous detection of carcinoembryonic antigen, alpha-fetoprotein, beta-human choriogonadotropin, and carcinoma antigen 125 in clinical serum samples with concentrations up to 188 microg/L, 250 microg/L, 266 IU/L, and 334 kIU/L, respectively. The detection limits (limits of the blank, mean of blank plus 3 SD) were 1.1 micro/L, 1.7 microg/L, 1.2 IU/L, and 1.7 kIU/L. The inter- and intraassay imprecision (CVs) of the immunosensor arrays were <7.8% and <9.0%, respectively. The immunosensor arrays were stable for 28 days.
This newly constructed immunosensor array provides a simple, automated, simultaneous multianalyte immunoassay with high throughput, short analytical time, and sufficiently low detection limits for clinical application. This method offers the capability of miniaturizing the multianalyte detection device.
临床诊断中越来越需要自动化、便捷且高通量的多分析物检测。我们开发了一种一次性4×2阵列，用于对4种肿瘤标志物进行程序化同步安培免疫分析。
我们采用了丝网印刷技术、一步固定法和流动注射技术。我们将癌胚抗原、甲胎蛋白、β-人绒毛膜促性腺激素和癌抗原125作为模型分析物固定在氧化还原介质接枝、生物聚合物修饰的丝网印刷碳电极阵列中，以在竞争性免疫反应中捕获相应的辣根过氧化物酶标记抗体。同步多分析物免疫分析自动进行，以安培法监测介质催化的对过氧化氢的酶促反应，该反应与样品中分析物的浓度成比例降低。
多分析物免疫传感器阵列的通量为每小时60个样品，能够同时检测临床血清样品中的癌胚抗原、甲胎蛋白、β-人绒毛膜促性腺激素和癌抗原125，其浓度分别高达188μg/L、250μg/L、266IU/L和334kIU/L。检测限（空白限，空白均值加3个标准差）分别为1.1μg/L、1.7μg/L、1.2IU/L和1.7kIU/L。免疫传感器阵列的批间和批内不精密度（CV）分别<7.8%和<9.0%。免疫传感器阵列在28天内稳定。
这种新构建的免疫传感器阵列提供了一种简单、自动化、同步的多分析物免疫分析方法，具有高通量、分析时间短和临床应用检测限足够低的特点。该方法具有使多分析物检测设备小型化的能力。
