Part:BBa_K5073010:Design
GP3C scFv
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
During the design of the GPC3 scFv sequence, several key considerations were taken into account:
Linker length and flexibility: The peptide linker connecting the VH and VL regions needed to be of sufficient length and flexibility to allow proper folding and maintain the scFv's binding affinity. Typically, a (Gly4Ser)3 linker is used to ensure flexibility without disrupting function.
Codon optimization: The sequence was optimized for expression in the intended host cells, such as mammalian cells (e.g., HEK-293T or CAR-T cells), to ensure efficient transcription and translation.
Protein folding and stability: Proper folding of the scFv was critical to maintain its binding specificity. Care was taken to avoid sequences that could lead to aggregation or misfolding.
Specificity and affinity: The design aimed to retain the high specificity and affinity of the original antibody for the GPC3 antigen while ensuring compatibility with downstream applications, such as CAR-T cell therapy.
Immunogenicity: Minimizing potential immunogenic regions was considered to reduce unwanted immune responses when used in therapeutic settings.
Source
The GPC3 scFv part is derived from the variable regions of an antibody that specifically targets the glypican-3 (GPC3) antigen. The sequence for this scFv was originally obtained through antibody screening techniques, such as phage display or hybridoma technology, designed to isolate antibodies against GPC3, a protein commonly overexpressed in hepatocellular carcinoma (HCC). The heavy and light chain variable regions (VH and VL) were genetically engineered and linked via a flexible peptide linker to create the single-chain variable fragment (scFv).