用于快速灵敏测定毒力决定因素的适配体固定化表面等离子体共振生物传感器

Aptamer-Immobilized Surface Plasmon Resonance Biosensor for Rapid and Sensitive Determination of Virulence Determinant.

作者信息

Song Myeong-Sub, Sekhon Simranjeet Singh, Shin Woo-Ri, Rhee Sung-Keun, Ko Jung Ho, Kim Sang Yong, Min Jiho, Ahn Ji-Young, Kim Yang-Hoon

机构信息

School of Biological Sciences, Chungbuk National University, 1 Chungdae-Ro, Seowon-Gu, Cheongju 28644, South Korea.

College of Veterinary Medicine, Western University of Health Sciences, 309 E Second Street, Pomona CA 91766, USA.

出版信息

J Nanosci Nanotechnol. 2018 May 1;18(5):3095-3101. doi: 10.1166/jnn.2018.14697.

DOI:10.1166/jnn.2018.14697

PMID:29442808

Abstract

Shigella sonnei isolate invasion plasmid antigen protein, IpaH, was successfully expressed in recombinant overexpression bacterial system. The soluble expression IpaH was enhanced with molecular chaperon co-expressed environment. Specific aptamer IpaH17 was isolated through the SELEX process and showed fM binding affinity. IpaH17-SPR biosensor platform was involved to verify the binding sensitivity and specificity. The IpaH concentration dependent IpaH17-SPR sensor response was highly linear with a linear regression constant of 99.4% in the range between 0 and 100 ng/mL. In addition, S. sonnei revealed the specific RU value and detected in a real-time manner within 1 hour. Our study indicated that IpaH17-SPR sensor can allow for rapid, sensitive and specific determination of Shigella sonnei virulent factor.

摘要

宋内志贺氏菌分离株的侵袭质粒抗原蛋白IpaH在重组过表达细菌系统中成功表达。在分子伴侣共表达环境下，IpaH的可溶性表达得到增强。通过SELEX过程分离出特异性适体IpaH17，其显示出飞摩尔级的结合亲和力。采用IpaH17-SPR生物传感器平台验证结合敏感性和特异性。IpaH浓度依赖性的IpaH17-SPR传感器响应在0至100 ng/mL范围内呈高度线性，线性回归常数为99.4%。此外，宋内志贺氏菌显示出特定的响应单位值，并在1小时内实现实时检测。我们的研究表明，IpaH17-SPR传感器能够快速、灵敏且特异地测定宋内志贺氏菌的毒力因子。