[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.
Si
B
B
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100850.
Yang
P
P
Qin
E
E
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics; D003001:Cloning, Molecular; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005803:Genes, Immunoglobulin / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D057127:Single-Chain Antibodies
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C114805:scFv fragment SW1