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

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 t...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d9eb0232ac50409ca353f40dbf298c2c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:d9eb0232ac50409ca353f40dbf298c2c
record_format	dspace
spelling	oai:doaj.org-article:d9eb0232ac50409ca353f40dbf298c2c2021-12-02T11:52:33ZPoly-protein G-expressing bacteria enhance the sensitivity of immunoassays10.1038/s41598-017-01022-w2045-2322https://doaj.org/article/d9eb0232ac50409ca353f40dbf298c2c2017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01022-whttps://doaj.org/toc/2045-2322Abstract 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.Wen-Rui HaoMichael ChenYi-Jou ChenYu-Cheng SuChiu-Min ChengHsiang-Yin HsuehAn-Pei KaoYuan-Chin HsiehJohny ChangMing-Yang TsengKuo-Hsiang ChuangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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 Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
description	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.
format	article
author	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
author_facet	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
author_sort	Wen-Rui Hao
title	Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
title_short	Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
title_full	Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
title_fullStr	Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
title_full_unstemmed	Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays
title_sort	poly-protein g-expressing bacteria enhance the sensitivity of immunoassays
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/d9eb0232ac50409ca353f40dbf298c2c
work_keys_str_mv	AT wenruihao polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT michaelchen polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT yijouchen polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT yuchengsu polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT chiumincheng polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT hsiangyinhsueh polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT anpeikao polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT yuanchinhsieh polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT johnychang polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT mingyangtseng polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays AT kuohsiangchuang polyproteingexpressingbacteriaenhancethesensitivityofimmunoassays
_version_	1718395014728384512

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

Ejemplares similares