A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
| Autor: | Zhong Wang, Yanlan Wang, Qing Li, Xiaoqiong Wu, Jiayu Liu, Haitao Pan, Jieyu Xing |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cancer Research General Chemical Engineering medicine.disease_cause Epitope General Biochemistry Genetics and Molecular Biology law.invention 03 medical and health sciences Immune system Glypicans law Antibodies Bispecific medicine Humans Cytotoxicity Escherichia coli biology General Immunology and Microbiology Chemistry General Neuroscience Molecular biology Fragment crystallizable region 030104 developmental biology Single-domain antibody Recombinant DNA biology.protein Antibody |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| Popis: | This protocol describes the construction and functional studies of a bispecific antibody (bsAb), GPC3-S-Fab. bsAbs can recognize two different epitopes through their two different arms. bsAbs have been actively studied for their ability to directly recruit immune cells to kill tumor cells. Currently, the majority of bsAbs are produced in the form of recombinant proteins, either as Fc-containing bsAbs or as smaller bsAb derivatives without the Fc region. In this study, GPC3-S-Fab, an antibody fragment (Fab) based bispecific antibody, was designed by linking the Fab of anti-GPC3 antibody GC33 with an anti-CD16 single domain antibody. The GPC3-S-Fab can be expressed in Escherichia coli and purified by two affinity chromatographies. The purified GPC3-S-Fab can specifically bind to and kill GPC3 positive liver cancer cells by recruiting natural killer cells, suggesting a potential application of GPC3-S-Fab in liver cancer therapy. |
| Databáze: | OpenAIRE |
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