两种基于阵列 SPRi 的用于检测血浆中 VEGF-R2 的生物传感器。

Two Biosensors for the Determination of VEGF-R2 in Plasma by Array SPRi.

机构信息

Faculty of Chemistry, Bioanalysis Laboratory, University of Bialystok, Ciolkowskiego 1K, 15-245 Bialystok, Poland.

Doctoral School of Exact and Natural Science, Faculty of Chemistry, University of Bialystok, Ciolkowskiego 1K, 15-245 Bialystok, Poland.

出版信息

Molecules. 2022 Dec 24;28(1):155. doi: 10.3390/molecules28010155.

DOI:10.3390/molecules28010155

PMID:36615347

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9822109/

Abstract

Vascular endothelial growth factor receptor 2 (VEGF-R2) is a marker of angiogenesis and metastasis of cancer. Two biosensors for the determination of VEGF-R2 in plasma have been developed. One of them is based on a pure gold chip, and the other on a silver/gold bimetallic chip; both have the receptor, monoclonal rabbit antibody specific for human VEGF-R2, attached to the chip via a cysteamine linker. The biosensor with the gold chip exhibits linearity of the analytical signal between 0.03 and 2 ng/mL, a precision of 1.4% and recovery between 99% and 102%. The biosensor with the bimetallic chip exhibits linearity between 0.03 and 1 ng/mL, a precision of 2.2% and recovery between 99% and 103%. Both biosensors tolerate a 1:100 excess of VEGF, VEGF-R1 and VEGF-R3. Both biosensors were validated by parallel determination of VEGF-R2 in 27 different plasma samples using the ELISA immunosensor assay, with very good agreement of the results. Thermodynamic parameters of the interaction of VEGF-R2 with the antibody were determined by QCM (Quartz Crystal Microbalance) and SPRi (Surface Plasmon Resonance imaging) measurements.

摘要

血管内皮生长因子受体 2（VEGF-R2）是癌症血管生成和转移的标志物。已经开发出两种用于测定血浆中 VEGF-R2 的生物传感器。其中一个基于纯金芯片，另一个基于银/金双金属芯片；两者都在芯片上附有受体，即针对人 VEGF-R2 的单克隆兔抗体，通过半胱氨酸接头连接到芯片上。金芯片生物传感器在 0.03 至 2ng/mL 之间表现出分析信号的线性，精度为 1.4%，回收率在 99%至 102%之间。双金属芯片生物传感器在 0.03 至 1ng/mL 之间表现出线性，精度为 2.2%，回收率在 99%至 103%之间。两种生物传感器都能耐受 VEGF、VEGF-R1 和 VEGF-R3 的 1:100 过量。两种生物传感器都通过使用 ELISA 免疫传感器测定法在 27 个不同的血浆样本中平行测定 VEGF-R2 进行了验证，结果非常吻合。通过 QCM（石英晶体微天平）和 SPRi（表面等离子体共振成像）测量确定了 VEGF-R2 与抗体相互作用的热力学参数。