Preparation of anti-GPC3 single chain antibody for targeted detection of hepatocellular carcinoma.

Xinchu Xiang; Xue Liu; Yaokai HU; Chengjun Zhao; Wenxin Luo

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Preparation of anti-GPC3 single chain antibody for targeted detection of hepatocellular carcinoma.

Author: Xinchu XIANG ¹ ; Xue LIU ¹ ; Yaokai HU ¹ ; Chengjun ZHAO ¹ ; Wenxin LUO ¹
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1. State key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian, China.
Publication Type:Journal Article
Keywords: GPC3; HCC; scFv; surgical navigation; targeted therapy
MeSH: Animals; Antibodies, Monoclonal; Carcinoma, Hepatocellular/genetics*; Glypicans/genetics*; Liver Neoplasms/diagnosis*; Mice
From: Chinese Journal of Biotechnology 2020;36(12):2860-2867
CountryChina
Language:Chinese
Abstract: Glypican-3 (GPC3) is a key member of Glypican family and plays an important role in the development, angiogenesis and metastasis of hepatocellular carcinoma (HCC). Most HCC overexpresses GPC3, but GPC3 is hardly detected in normal adult liver and benign liver lesions, so it is regarded as a highly specific diagnostic marker and an ideal therapeutic target for HCC. In this study, we cloned the heavy and light chain variable region gene from the monoclonal antibody targeted to GPC3 screened in the previous stage, and linked it with a segment of flexible peptide (Linker) to obtain the single chain antibody against GPC3. The single chain antibody gene was cloned into vector for prokaryotic expression and purified to obtain high purity protein. Detection shows that the single-chain antibody produced by us has the same binding activity with the full-length antibody, and can accurately target the tumor site of Huh7 tumor-bearing model mice after coupling Cy5.5 fluorescence, suggesting that the single-chain antibody has the potential to realize multi-directional liver cancer precise surgical navigation under the guidance of a probe.