Quantification of chromogranin A using a surface plasmon resonance-based biosensor
Quantification of chromogranin A using a surface plasmon resonance-based biosensor
复制标题
使用基于表面等离子共振的生物传感器对嗜铬粒蛋白 A 进行定量
DOI：
10.1039/d1ay00782c
复制
发表时间：
2021
影响因子：
3.1
通讯作者：
Gao Jinhang
中科院分区：
文献类型：
--
作者：
Xiao Yang;Tai Yang;Quan Xin;Zhao Chong;Liu Rui;Tong Huan;Huang Zhiyin;Tang Chengwei;Gao Jinhang
血液中的嗜铬粒蛋白A（CgA）水平是神经内分泌肿瘤和其他疾病的重要生物标志物。传统的CgA检测方法成本高、耗时长、重复性差。在这项研究中，表面等离子体共振（SPR），一个简单的和无标记的技术，被验证用于定量CgA。将CgA抗体（CgA-Ab）在最佳条件（pH 5.0，10 μg mL-1）下固定在CM 5传感器芯片（CgA-Ab-CM 5）上。接着，通过CgA-Ab-CM 5测量不同浓度的CgA。优化结合和再生条件并用于每次测量。选择240 s的结合时间和30 μL min−1的流速作为最佳结合条件。pH值为1.75时再生效果最好。与酶联免疫吸附测定（ELISA）的检测范围23.4-187 ng mL−1相比，基于响应单位（RU）检测的线性范围为0.2-187 ng mL−1，显示SPR的高灵敏度和可靠性。最后，通过连续结合-再生循环300次来评估CgA-Ab-CM 5芯片的再现性。总之，基于SPR的CgA-Ab-CM 5芯片是以实时方式定量CgA水平的灵敏且可重复的方法。
The chromogranin A (CgA) level in the blood is an important biomarker for neuroendocrine tumors and other diseases. Traditional methods for detecting CgA are expensive and time-consuming with low reproducibility. In this study, surface plasmon resonance (SPR), a simple and label-free technique, was validated for quantifying CgA. CgA antibody (CgA-Ab) was immobilized on the CM5 sensor chip (CgA-Ab-CM5) at optimal conditions (pH 5.0, 10 μg mL−1). Next, different concentrations of CgA were measured by CgA-Ab-CM5. The binding and regeneration conditions were optimized and used in each measurement. A binding time of 240 s, and flow rate of 30 μL min−1 were chosen as the optimal binding conditions. A pH of 1.75 was the optimal regeneration condition. Compared to the detection range of 23.4–187 ng mL−1 for enzyme-linked immunosorbent assay (ELISA), a linear range of 0.2–187 ng mL−1 was detected based on the response unit (RU), showing high sensitivity and reliability of SPR. Finally, the reproducibility of the CgA-Ab-CM5 chip was accessed by consecutive binding-regeneration cycles for 300 times. In conclusion, the SPR-based CgA-Ab-CM5 chip is a sensitive and reproducible method for quantifying CgA levels in a real-time manner.