Expression and characterization of single-chain variable fragment antibody against staphylococcal enterotoxin A in Escherichia coli.

Abstract	The staphylococcal enterotoxins (SEs) are potent gastrointestinal exotoxins synthesized by Staphylococcus aureus, which is responsible for various diseases including septicemia, food poisoning, and toxic shock syndrome, as well as bovine mastitis. Among them, staphylococcal enterotoxin A (SEA) is one of the most commonly present serotypes in staphylococcal food poisoning cases. In this study, the stable hybridoma 3C12 producing anti-SEA monoclonal antibody was established with an equilibrium dissociation constant (KD) of 1.48 × 10(-8) mol·L(-1), its ScFv-coding genes were obtained and then the anti-SEA single chain variable fragment (ScFv) protein was expressed in Escherichia coli. Characterization of the expressed target ScFv protein was analyzed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis, Western blot, and enzyme-linked immunosorbent assay. The results demonstrated that the recombinant anti-SEA ScFv protein retained a specific binding activity for SEA, and the KD value of the soluble ScFv was about 3.75 × 10(-7) mol·L(-1). The overall yield of bioactive anti-SEA ScFv in E. coli flask culture was more than 10 mg·L(-1).
Authors	Weifeng Chen, Li Hu, Aiping Liu, Jinquan Li, Fusheng Chen, Xiaohong Wang
Journal	Canadian journal of microbiology (Can J Microbiol) Vol. 60 Issue 11 Pg. 737-43 (Nov 2014) ISSN: 1480-3275 [Electronic] Canada
PMID	25322256 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Enterotoxins Recombinant Proteins Single-Chain Antibodies enterotoxin A, Staphylococcal
Topics	Amino Acid Sequence Antibody Affinity Antibody Specificity Base Sequence Blotting, Western Electrophoresis, Polyacrylamide Gel Enterotoxins (immunology) Enzyme-Linked Immunosorbent Assay Escherichia coli (genetics) Genes, Immunoglobulin Heavy Chain Genes, Immunoglobulin Light Chain Hybridomas Molecular Sequence Data Recombinant Proteins (immunology) Single-Chain Antibodies (biosynthesis, chemistry, genetics, immunology) Staphylococcus aureus (immunology)

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