Clone and expression of adiponectin single chain antibody gene

Objective To construct and express the single chain antibody(scFv)in E.coli JM107 by cloning the variable region genes from habridoma against adiponectin.Methods Total RNA was extracted from hybridoma cell line secreting monoclonal antibodies(McAbs)against adiponectin,and the cDNAs of VL and VH were amplified by RT PCR.The products were fused by a short peptide linker and inserted into pUC19 vector and expressed in E.coli JM109,then their sequences were analyzed.Results The VH gene contained 350bp encoding 116 amino acid residues.The VL gene contained 340bp encoding 113 amino acid residues.Restriction endonuclease digestion and DNA sequencing proved that the scFv contained 710bp that was constructed correctly.ELISA results showed that scFv was successfully expressed in E.coli JM109 and the expression protein had specific antigen binding activity.Conclusion Anti-human adiponectin ScFv is successfully constructed for the use in clinical studies.