Construction of non-covalent single-chain Fv dimers for hepatocellular carcinoma and their biological functions.

ABSTRACT:

Aim

To create new diabodies with improved binding activity to antigen of the variable light - variable heavy (VH-VL) oriented single-chain Fv dimers genes (scFv).

Methods

The linker between VH and VL genes was shortened to 3-5 amino acid residues and cloned into the vector pCANTAB5E. The recombinant plasmids were transformed into TG1 cells and sequenced. The positive transformed cells were infected by M13K07 helper phage to form human recombinant phage antibodies. Expressed products were identified by SDS-PAGE, Western blotting, size exclusion gel chromatography (SEC), ELISA and immunohistochemistry.

Results

Three scFv (scFv-3, scFv-4, scFv-5) were constructed successfully with binding ability to hepatocellular carcinoma 3.5-6 fold greater than their parental scFv. Th

SUBMITTER: Bie CQ 

PROVIDER: S-EPMC2998965 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature