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Expression and bioactivity identification of soluble MG7 scFv
Zhao-Cai Yu1, Jie Ding, Bo-Rong Pan
1Department of Gastroenterology, Xijing Hospital, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China.
World Journal of Gastroenterology
|February 8, 2002
Summary
Researchers developed soluble MG7 single-chain variable fragment (scFv) with strong antigen-binding affinity for gastric cancer gene therapy. This antibody fragment shows potential as a targeting mediator in future therapeutic applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Gastric cancer remains a significant global health challenge.
- Targeted gene therapy offers a promising approach for cancer treatment.
- The MG7 antibody targets gastric cancer cells, but its application requires further development.
Purpose of the Study:
- To characterize the molecular mass and bioactivity of the MG7 single-chain variable fragment (scFv).
- To evaluate the potential of MG7 scFv as a targeting mediator in gastric cancer gene therapy.
Main Methods:
- Recombinant phage display technology was used to screen and isolate MG7 scFv clones.
- Escherichia coli (E. coli) strains (TG1 and HB2151) were utilized for gene cloning and protein expression.
- Enzyme-linked immunosorbent assay (ELISA), immunodotting, and Western blot were employed to assess antigen-binding affinity, yield, and molecular mass.
- DNA sequencing was performed to analyze the nucleotide sequence of the MG7 scFv.
Main Results:
- Two positive recombinant phage clones containing the scFv gene were identified.
- MG7 scFv demonstrated strong antigen-binding affinity via ELISA.
- Transfected E. coli HB2151 successfully produced soluble MG7 scFv with high yield upon IPTG induction.
- Western blot analysis confirmed the molecular mass of MG7 scFv to be approximately 30 kDa.
Conclusions:
- The soluble MG7 scFv was successfully developed.
- The characterized MG7 scFv exhibits strong antigen-binding capabilities.
- This engineered antibody fragment holds potential for targeted delivery in gastric cancer gene therapy.