Hypersensitive immunoassay by using Escherichia coli outer membrane with autodisplayed Z-domains

详细信息    查看全文

• 作者：Min Park ; Joachim Jose ; Jae-Chul Pyun
• 关键词：Immunoassay ; Z-domain ; Orientation control ; C-reactive protein ; Autodisplay ; Outer membrane
• 刊名：Enzyme and Microbial Technology
• 出版年：2010
• 出版时间：5 March 2010
• 年：2010
• 卷：46
• 期：3-4
• 页码：309-314
• 全文大小：710 K

文摘

In this work, the Z-domain of protein A was autodisplayed on the surface of the Escherichia coli outer membrane as a fusion protein of AIDA-1, and then layered on a microplate in order to develop a hypersensitive immunoassay. By using microplates with controlled hydrophilicity the formation of outer membrane layer was carried out, and the driving force for the layer formation was proven to be a hydrophobic interactions. Through the orientation control of the immobilized antibodies by using autodisplayed Z-domains, the limit of detection (LOD) and the sensitivity of this new immunoassay were determined to have improved as much as 30-fold compared to the conventional ELISA. The applicability of the new immunoassay for medical diagnosis was demonstrated by the detection of C-reactive protein (CRP) which is known to be a biomarker protein for inflammatory diseases.