Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Wen Rui Hao, Michael Chen, Yi Jou Chen, Yu-Cheng Su, Chiu Min Cheng, Hsiang Yin Hsueh, An Pei Kao, Yuan Chin Hsieh, Johny Chang, Ming Yang Tseng, Kuo Hsiang Chuang*

*Corresponding author for this work

研究成果: Article同行評審

4 引文 斯高帕斯(Scopus)

摘要

The sensitivities of solid-phase immunoassays are limited by the quantity of detection antibodies bound to their antigens on the solid phase. Here, we developed a poly-protein G-expressing bacterium as an antibody-trapping microparticle to enhance the signals of immunoassays by increasing the accumulation of detection antibodies on the given antigen. Eight tandemly repeated fragment crystallisable (Fc) binding domains of protein G were stably expressed on the surface of Escherichia coli BL21 cells (termed BL21/8G). BL21/8G cells showed a higher avidity for trapping antibodies on their surface than monomeric protein G-expressing BL21 (BL21/1G) cells did. In the sandwich enzyme-linked immunosorbent assay (ELISA), simply mixing the detection antibody with BL21/8G provided a detection limit of 6 pg/mL for human interferon-α (IFN-α) and a limit of 30 pg/mL for polyethylene glycol (PEG)-conjugated IFN-α (Pegasys), which are better than that of the traditional ELISA (30 pg/mL for IFN-α and 100 pg/mL for Pegasys). Moreover, the sensitivity of the Western blot for low-abundance Pegasys (0.4 ng/well) was increased by 25 folds upon mixing of an anti-PEG antibody with BL21/8G cells. By simply being mixed with a detection antibody, the poly-protein G-expressing bacteria can provide a new method to sensitively detect low-abundance target molecules in solid-phase immunoassays.

原文English
文章編號989
期刊Scientific reports
7
發行號1
DOIs
出版狀態Published - 1 十二月 2017

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