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

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

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3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number989
JournalScientific Reports
Volume7
Issue number1
DOIs
Publication statusPublished - Dec 1 2017

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Bacteria
Antibodies
Proteins
Interferons
Immunosorbents
Assays
Antigens
Monomeric GTP-Binding Proteins
Enzymes
Escherichia coli
Molecules
peginterferon alfa-2a

ASJC Scopus subject areas

  • General

Cite this

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays. / Hao, Wen Rui; Chen, Michael; Chen, Yi Jou; Su, Cheng Yu; Cheng, Chiu Min; Hsueh, Hsiang Yin; Kao, An Pei; Hsieh, Yuan Chin; Chang, Johny; Tseng, Ming Yang; Chuang, Kuo Hsiang.

In: Scientific Reports, Vol. 7, No. 1, 989, 01.12.2017.

Research output: Contribution to journalArticle

Hao, WR, Chen, M, Chen, YJ, Su, CY, Cheng, CM, Hsueh, HY, Kao, AP, Hsieh, YC, Chang, J, Tseng, MY & Chuang, KH 2017, 'Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays', Scientific Reports, vol. 7, no. 1, 989. https://doi.org/10.1038/s41598-017-01022-w
Hao, Wen Rui ; Chen, Michael ; Chen, Yi Jou ; Su, Cheng Yu ; Cheng, Chiu Min ; Hsueh, Hsiang Yin ; Kao, An Pei ; Hsieh, Yuan Chin ; Chang, Johny ; Tseng, Ming Yang ; Chuang, Kuo Hsiang. / Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays. In: Scientific Reports. 2017 ; Vol. 7, No. 1.
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