Simply Mixing Poly Protein G with Detection Antibodies Enhances the Detection Limit and Sensitivity of Immunoassays

Yi Jou Chen, Michael Chen, Tian Lu Cheng, Steve R. Roffler, Shyr Yi Lin, Hui Lan Hsu, Chang Hung Wang, Che Yi Chen, An Pei Kao, Jing Jy Cheng, Kuo Hsiang Chuang

研究成果: 雜誌貢獻文章

摘要

An insufficient amount of detection antibodies bound to their antigens usually limits the sensitivity of immunoassays. Here, we describe a simple method to improve the detection limit and sensitivity of various immunoassays by mixing detection antibodies with a soluble poly protein G (named 8pG). 8pG was developed by fusing eight repeated fragment crystallizable (Fc) binding domains of streptococcal protein G to a linear polymer. Simply mixing detection antibodies with 8pG to form an antibody/8pG complex largely increased the accumulation of detection antibody to target molecules, which dramatically enhanced the sensitivity in direct ELISA, sandwich ELISA, Western blot, and flow cytometry systems, separately. The detection limit of Western blot for low-abundance PEGylated interferon (Pegasys) and recombinant human CTLA4 (rhCTLA4) improved by at least 13-fold and 31-fold, respectively, upon mixing detection antibodies with 8pG. Moreover, the nanoscale size of the antibody/8pG complex did not influence the granularity and dimension of target cells in the flow cytometry system. Collectively, we provide a quick and easy-to-operate method to make various immunoassays to sensitively detect low-abundance target molecules by just mixing their detection antibodies with 8pG.

原文英語
期刊Analytical Chemistry
DOIs
出版狀態已發佈 - 一月 1 2019

指紋

Antibodies
Proteins
Flow cytometry
Molecules
Interferons
Polymers
Antigens

ASJC Scopus subject areas

  • Analytical Chemistry

引用此文

Simply Mixing Poly Protein G with Detection Antibodies Enhances the Detection Limit and Sensitivity of Immunoassays. / Chen, Yi Jou; Chen, Michael; Cheng, Tian Lu; Roffler, Steve R.; Lin, Shyr Yi; Hsu, Hui Lan; Wang, Chang Hung; Chen, Che Yi; Kao, An Pei; Cheng, Jing Jy; Chuang, Kuo Hsiang.

於: Analytical Chemistry, 01.01.2019.

研究成果: 雜誌貢獻文章

Chen, Yi Jou ; Chen, Michael ; Cheng, Tian Lu ; Roffler, Steve R. ; Lin, Shyr Yi ; Hsu, Hui Lan ; Wang, Chang Hung ; Chen, Che Yi ; Kao, An Pei ; Cheng, Jing Jy ; Chuang, Kuo Hsiang. / Simply Mixing Poly Protein G with Detection Antibodies Enhances the Detection Limit and Sensitivity of Immunoassays. 於: Analytical Chemistry. 2019.
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abstract = "An insufficient amount of detection antibodies bound to their antigens usually limits the sensitivity of immunoassays. Here, we describe a simple method to improve the detection limit and sensitivity of various immunoassays by mixing detection antibodies with a soluble poly protein G (named 8pG). 8pG was developed by fusing eight repeated fragment crystallizable (Fc) binding domains of streptococcal protein G to a linear polymer. Simply mixing detection antibodies with 8pG to form an antibody/8pG complex largely increased the accumulation of detection antibody to target molecules, which dramatically enhanced the sensitivity in direct ELISA, sandwich ELISA, Western blot, and flow cytometry systems, separately. The detection limit of Western blot for low-abundance PEGylated interferon (Pegasys) and recombinant human CTLA4 (rhCTLA4) improved by at least 13-fold and 31-fold, respectively, upon mixing detection antibodies with 8pG. Moreover, the nanoscale size of the antibody/8pG complex did not influence the granularity and dimension of target cells in the flow cytometry system. Collectively, we provide a quick and easy-to-operate method to make various immunoassays to sensitively detect low-abundance target molecules by just mixing their detection antibodies with 8pG.",
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