Full
Outline
To construct bispecific fusion protein from Glypican-3 and NKG2D, and to primarily evaluate the cytolytic properties against human hepatocellular carcinoma cell line HepG2 in vitro.
Using genetic engineering technology, bispecific fusion protein FabGC33-MICA (human MHC class Ⅰ chain-related A, FabGM) expression vector was obtained by fusion the extracellular domain of MICA to heavy chain's C-terminal of the Fab form of GC33, and was used to transfect FreeStyleTM 293-F cells. FabGM fusion protein was purified from the cell supernatant by affinity chromatography. Octet QKe system (ForteBio) was applied to detect FabGM's affinity for its targets. Flow cytometry analysis was used to assess the avidity of FabGM fusion protein in HepG2 cells. The cytotoxicity of FabGM against HepG2 was monitored by IncuCyte Live Cell Imaging System. The secretion of interferon-γ (IFN-γ) by PBMCs was detected by ELISA assay.
FabGM was successfully purified and confirmed selectively combined with GPC3 and NKG2D. The SDS-PAGE assay confirmed the predicted molecular. The Kd values of FabGM to target molecular GPC3 and NKG2D was 1.14 and 561 nmol·L-1 respectively. FabGM effectively mediated the targeted killing effect of PBMCs against HepG2. ELISA assay showed that IFN-γ level was significantly increased during the killing process compared with negative control.
Bispecific fusion protein FabGM effectively mediated PBMCs to kill HepG2 cells in vitro, providing experimental evidence for FabGM to become a candidate drug for hepatocellular carcinoma treatment.
30
6
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
关闭全屏
No. 86 Xueyuan South Road, Haidian District, Beijing
010-62662699(Operation and Technical Support)
qkjq@cast.org.cn
Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House
