Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma

Lijie Xia, Qiao Teng, Qi Chen, Fuchun Zhang Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, People’s Republic of ChinaCorrespondence: Lijie XiaXinjiang Key Laboratory of Biological Resource...

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Autores principales:	Xia L, Teng Q, Chen Q, Zhang F
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	gpc3 phage display library selection nanobody Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/86522d355653406db1f2e1d115f2d0a0
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spelling	oai:doaj.org-article:86522d355653406db1f2e1d115f2d0a02021-12-02T08:15:40ZPreparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma1178-2013https://doaj.org/article/86522d355653406db1f2e1d115f2d0a02020-03-01T00:00:00Zhttps://www.dovepress.com/preparation-and-characterization-of-anti-gpc3-nanobody-against-hepatoc-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Lijie Xia, Qiao Teng, Qi Chen, Fuchun Zhang Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, People’s Republic of ChinaCorrespondence: Lijie XiaXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, 666 Shengli Road, Urumqi, Xinjiang 830046, People’s Republic of ChinaEmail xialijie1219@163.comBackground: Glypican-3 (GPC3) is a newly identified target molecule for the early diagnosis of hepatocellular carcinoma (HCC), while targeted inhibition of GPC3 signaling may help to control the proliferation and metastasis of HCC cells. The purpose of this study was to prepare the anti-GPC3 nanobody and to investigate the affinity of the anti-GPC3 nanobodies in vitro and the anticancer effects on hepatocellular carcinoma in vivo.Methods: To screen for unknown anti-GPC3 antibodies, we constructed an antibody phage display library. After three rounds of panning, positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA). Further, the nanobody fusion protein was expressed in E. coli BL21 cells and purified by affinity chromatography. Competitive ELISA and flow cytometry were conducted to confirm the affinity of the anti-GPC3 nanobodies in vitro. The antitumor effects of VHHGPC3 were assessed in vivo.Results: The results showed that the nanobody VHHGPC3 had specific high-affinity binding to His-GPC3 antigen. Moreover, VHHGPC3 exhibited specific binding to commercial human GPC3 and recognized the surface GPC3 protein of the hepatoma cell line HepG2. Importantly, in vivo study showed that GPC3 nanobody suppresses the growth of HepG2 and improves the survival rate of tumor mice.Discussion: In summary, our new anti-GPC3 nanobody suggests a strong application potential for targeted therapy of liver cancer.Keywords: GPC3, phage display library, selection, nanobodyXia LTeng QChen QZhang FDove Medical Pressarticlegpc3phage display libraryselectionnanobodyMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 15, Pp 2197-2205 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	gpc3 phage display library selection nanobody Medicine (General) R5-920
spellingShingle	gpc3 phage display library selection nanobody Medicine (General) R5-920 Xia L Teng Q Chen Q Zhang F Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
description	Lijie Xia, Qiao Teng, Qi Chen, Fuchun Zhang Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, People’s Republic of ChinaCorrespondence: Lijie XiaXinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, 666 Shengli Road, Urumqi, Xinjiang 830046, People’s Republic of ChinaEmail xialijie1219@163.comBackground: Glypican-3 (GPC3) is a newly identified target molecule for the early diagnosis of hepatocellular carcinoma (HCC), while targeted inhibition of GPC3 signaling may help to control the proliferation and metastasis of HCC cells. The purpose of this study was to prepare the anti-GPC3 nanobody and to investigate the affinity of the anti-GPC3 nanobodies in vitro and the anticancer effects on hepatocellular carcinoma in vivo.Methods: To screen for unknown anti-GPC3 antibodies, we constructed an antibody phage display library. After three rounds of panning, positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA). Further, the nanobody fusion protein was expressed in E. coli BL21 cells and purified by affinity chromatography. Competitive ELISA and flow cytometry were conducted to confirm the affinity of the anti-GPC3 nanobodies in vitro. The antitumor effects of VHHGPC3 were assessed in vivo.Results: The results showed that the nanobody VHHGPC3 had specific high-affinity binding to His-GPC3 antigen. Moreover, VHHGPC3 exhibited specific binding to commercial human GPC3 and recognized the surface GPC3 protein of the hepatoma cell line HepG2. Importantly, in vivo study showed that GPC3 nanobody suppresses the growth of HepG2 and improves the survival rate of tumor mice.Discussion: In summary, our new anti-GPC3 nanobody suggests a strong application potential for targeted therapy of liver cancer.Keywords: GPC3, phage display library, selection, nanobody
format	article
author	Xia L Teng Q Chen Q Zhang F
author_facet	Xia L Teng Q Chen Q Zhang F
author_sort	Xia L
title	Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
title_short	Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
title_full	Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
title_fullStr	Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
title_full_unstemmed	Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma
title_sort	preparation and characterization of anti-gpc3 nanobody against hepatocellular carcinoma
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/86522d355653406db1f2e1d115f2d0a0
work_keys_str_mv	AT xial preparationandcharacterizationofantigpc3nanobodyagainsthepatocellularcarcinoma AT tengq preparationandcharacterizationofantigpc3nanobodyagainsthepatocellularcarcinoma AT chenq preparationandcharacterizationofantigpc3nanobodyagainsthepatocellularcarcinoma AT zhangf preparationandcharacterizationofantigpc3nanobodyagainsthepatocellularcarcinoma
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Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma

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