[Soluble expression and identification of expressed single chain antibody fragment 3D3 against dengue virus type 3]

Hybridoma secreting 3D3 monoclonal antibody against dengue virus type 3 was used. The isolated mRNA was reverse-transcribed into cDNA for amplifying the heavy and light chain variable domains genes. The amplified light and heavy chain variable domains' genes were connected by flexible linker to from a single chain fragment (ScFv) gene of 750 bp, which was cloned into phage pCANTAB5E vector using the recombination phage antibody system. pCANTAB5E vector transformed E coli HB2151. Soluble single chain antibody fragment was expressed both in the bacterial supernatant and cell periplasm under the induction of IPTG. The expressed single chain antibody fragment has a molecular weight of about 28 kD detected by SDS-PAGE and can react with dengue virus type 3 by immunofluorescence test.