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Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3

Glypican-3 (GPC3) is a promising new marker for hepatocellular carcinoma, but the reported values for serum GPC3 differ markedly between currently available kits. Here we isolated Affimer non-antibody binding proteins against GPC3 by phage display and developed a new sandwich chemiluminescence immun...

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Autores principales: Xie, Chunmei, Tiede, Christian, Zhang, Xuanyi, Wang, Congrong, Li, Zhixiong, Xu, Xiao, McPherson, Michael J., Tomlinson, Darren C., Xu, Weiwen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5575301/
https://www.ncbi.nlm.nih.gov/pubmed/28852111
http://dx.doi.org/10.1038/s41598-017-10083-w
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author Xie, Chunmei
Tiede, Christian
Zhang, Xuanyi
Wang, Congrong
Li, Zhixiong
Xu, Xiao
McPherson, Michael J.
Tomlinson, Darren C.
Xu, Weiwen
author_facet Xie, Chunmei
Tiede, Christian
Zhang, Xuanyi
Wang, Congrong
Li, Zhixiong
Xu, Xiao
McPherson, Michael J.
Tomlinson, Darren C.
Xu, Weiwen
author_sort Xie, Chunmei
collection PubMed
description Glypican-3 (GPC3) is a promising new marker for hepatocellular carcinoma, but the reported values for serum GPC3 differ markedly between currently available kits. Here we isolated Affimer non-antibody binding proteins against GPC3 by phage display and developed a new sandwich chemiluminescence immunoassay (CLIA) combining an Affimer with a monoclonal antibody (Affimer-MAb CLIA). The proposed CLIA assay demonstrated a wide linear range  0.03–600 ng/mL) with a good linear correlation coefficient (0.9999), a high detection limitation (0.03 ng/mL) and specificity (0–0.002%) for detection of GPC3. The accuracy, hook effect and stability were demonstrated to be satisfactory. The mean level of GPC3 in serum was higher (>8.5 fold, P < 0.001) in hepatocellular carcinoma patients compared to healthy and other liver disease individuals. A poor correlation (correlation coefficients ranged from −0.286 to 0.478) was observed through pairwise comparison within different kits. However, only this newly developed CLIA test showed high specificity and correlated with the “gold standard” GPC3-immunohistochemistry. This study indicates that Affimer-MAb CLIA can be used to generate a sensitive immunodiagnostic kit, which offers the potential for a highly specific clinically-relevant detection system.
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spelling pubmed-55753012017-09-01 Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3 Xie, Chunmei Tiede, Christian Zhang, Xuanyi Wang, Congrong Li, Zhixiong Xu, Xiao McPherson, Michael J. Tomlinson, Darren C. Xu, Weiwen Sci Rep Article Glypican-3 (GPC3) is a promising new marker for hepatocellular carcinoma, but the reported values for serum GPC3 differ markedly between currently available kits. Here we isolated Affimer non-antibody binding proteins against GPC3 by phage display and developed a new sandwich chemiluminescence immunoassay (CLIA) combining an Affimer with a monoclonal antibody (Affimer-MAb CLIA). The proposed CLIA assay demonstrated a wide linear range  0.03–600 ng/mL) with a good linear correlation coefficient (0.9999), a high detection limitation (0.03 ng/mL) and specificity (0–0.002%) for detection of GPC3. The accuracy, hook effect and stability were demonstrated to be satisfactory. The mean level of GPC3 in serum was higher (>8.5 fold, P < 0.001) in hepatocellular carcinoma patients compared to healthy and other liver disease individuals. A poor correlation (correlation coefficients ranged from −0.286 to 0.478) was observed through pairwise comparison within different kits. However, only this newly developed CLIA test showed high specificity and correlated with the “gold standard” GPC3-immunohistochemistry. This study indicates that Affimer-MAb CLIA can be used to generate a sensitive immunodiagnostic kit, which offers the potential for a highly specific clinically-relevant detection system. Nature Publishing Group UK 2017-08-29 /pmc/articles/PMC5575301/ /pubmed/28852111 http://dx.doi.org/10.1038/s41598-017-10083-w Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Xie, Chunmei
Tiede, Christian
Zhang, Xuanyi
Wang, Congrong
Li, Zhixiong
Xu, Xiao
McPherson, Michael J.
Tomlinson, Darren C.
Xu, Weiwen
Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title_full Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title_fullStr Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title_full_unstemmed Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title_short Development of an Affimer-antibody combined immunological diagnosis kit for glypican-3
title_sort development of an affimer-antibody combined immunological diagnosis kit for glypican-3
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5575301/
https://www.ncbi.nlm.nih.gov/pubmed/28852111
http://dx.doi.org/10.1038/s41598-017-10083-w
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