Part:BBa_K5073010

GP3C scFv

GPC3 scFv (single-chain variable fragment) is a genetically engineered antibody fragment designed to specifically recognize the glypican-3 (GPC3) antigen, which is overexpressed in hepatocellular carcinoma (HCC) and other cancers. This scFv consists of the variable regions of both the heavy and light chains (VH and VL) connected by a flexible linker. It is typically used in CAR-T cell therapy, where it functions as the antigen-binding domain of the chimeric antigen receptor (CAR), allowing T cells to specifically recognize and attack GPC3-positive cancer cells.

To use this part in your project, you can clone the GPC3 scFv gene into a suitable vector for expression in your system, typically in CAR-T cells or other immune cell-based therapies. It's essential to ensure that your expression system is optimized for proper folding and surface display of the scFv. This part can be integrated into various therapeutic or diagnostic platforms targeting GPC3-expressing tumors.

GPC3 scFv proof (single-chain variable fragment) Given that GPC3 is a tumor-associated antigen that is overexpressed in HCC but is almost absent in normal liver tissues and most other normal tissues, numerous GPC3 CAR-T projects have already entered the clinical trial stage, demonstrating its feasibility as a target [6]. Therefore, we chose GPC3 as a target for optimizing CAR-T therapy.

We referenced to studies that have entered the clinical stage and drew on their GPC3 scFv sequence design. At the beginning of the project, we performed molecular simulations of the interaction between the GPC3 protein and the GPC3 scFv sequence using AlphaFold3, which showed good binding ability. Subsequently, we further verified this finding through a series of wet experiments, which showed that GPC3 was able to successfully activate the CAR molecule, fully demonstrating the functional feasibility of the design.

We established a TIDE score model to simulate the response of GPC3 high-expressing liver cancer cells to immunosuppressants. The results showed that liver cancer cells with high GPC3 expression had higher TIDE scores, suggesting these cells may possess stronger immune escape ability. As a result, they may respond poorly to immune checkpoint inhibitors, suggesting that a higher intensity of T-cell stimulation may be required to overcome this resistance.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]