用于使用噬菌体展示抗体检测单核细胞增生李斯特菌的表面等离子体共振生物传感器。

SPR biosensor for the detection of L. monocytogenes using phage-displayed antibody.

作者信息

Nanduri Viswaprakash, Bhunia Arun K, Tu Shu-I, Paoli George C, Brewster Jeffrey D

机构信息

Center For Food Safety and Engineering, Department of Food Science, Purdue University, 745 Agriculture Mall Dr., West Lafayette, IN 47907, USA.

出版信息

Biosens Bioelectron. 2007 Sep 30;23(2):248-52. doi: 10.1016/j.bios.2007.04.007. Epub 2007 Apr 19.

DOI:10.1016/j.bios.2007.04.007

PMID:17512186

Abstract

Whole cells of Listeria monocytogenes were detected with a compact, surface plasmon resonance (SPR) sensor using a phage-displayed scFv antibody to the virulence factor actin polymerization protein (ActA) for biorecognition. Phage Lm P4:A8, expressing the scFv antibody fused to the pIII surface protein was immobilized to the sensor surface through physical adsorption. A locally constructed fluidics system was used to deliver solutions to the compact, two-channel SPREETA sensor. Specificity of the sensor was tested using common food-borne bacteria and a control phage, M13K07 lacking the scFv fusion on its coat protein. The detection limit for L. monocytogenes whole cells was estimated to be 2 x 10(6)cfu/ml. The sensor was also used to determine the dissociation constant (Kd) for the interaction of phage-displayed scFv and soluble ActA in solution as 4.5 nM.

摘要

利用一种针对毒力因子肌动蛋白聚合蛋白（ActA）的噬菌体展示单链抗体片段（scFv）进行生物识别，通过紧凑型表面等离子体共振（SPR）传感器检测单核细胞增生李斯特菌的全细胞。表达与pIII表面蛋白融合的scFv抗体的噬菌体Lm P4:A8通过物理吸附固定在传感器表面。使用本地构建的流体系统将溶液输送到紧凑型双通道SPREETA传感器。使用常见食源细菌和一种对照噬菌体M13K07（其外壳蛋白上缺乏scFv融合）测试传感器的特异性。单核细胞增生李斯特菌全细胞的检测限估计为2×10⁶cfu/ml。该传感器还用于确定溶液中噬菌体展示的scFv与可溶性ActA相互作用的解离常数（Kd）为4.5 nM。