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Abstract: Staphylococcal enterotoxin A (SEA) synthesized by Staphylococcus aureus is a foodborne and heat-stable toxin, which is a great threat to human health (Pexara et al., 2010). Highly sensitive antibodies are a key factor in the immunological detection of SEA, which is one of the most effective ways to detect SEA because of its accuracy, agility, and efficiency (Nouri et al., 2018). In this study, we constructed a tetravalent anti-SEA antibody gene by linking the tetramerization domain of human p53 to the C-terminus of the anti-SEA single-chain variable fragment (scFv), which was then transformed into Escherichia coli BL21 (DE3) for the production of a SEA-specific tetravalent antibody. Successful expression of the tetravalent antibody was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot. An indirect non-competitive enzyme-linked immunosorbent assay (ELISA) revealed that the tetravalent antibody exhibited SEA-specific binding activity. A sandwich ELISA demonstrated that compared to the scFv monomer, the tetravalent antibody was more sensitive in detecting SEA. Molecular docking analysis revealed that the SEA interacted with the scFv mainly on the opposite side of the residue linked to p53. Thus, this study indicated that genetically engineered tetramerization is a potential way to improve the sensitivity of SEA-specific scFv.

一种用于检测金黄色葡萄球菌A型肠毒素的四聚化单链抗体

[1]ChenWF, HuL, LiuAP, et al., 2014. Expression and characterization of single-chain variable fragment antibody against staphylococcal enterotoxin A in Escherichia coli. Can J Microbiol, 60(11):737-743.

[2]DasguptaA, 2019. Biotin and Other Interferences in Immunoassays: A Concise Guide. Elsevier, St. Louis.

[3]LiangB, ZhangYX, LiuAP, et al., 2011. Production of a monoclonal antibody by simultaneous immunization of staphylococcal enterotoxin A and B. Appl Biochem Biotechnol, 164(6):831-840.

[4]LiuJL, AndersonGP, GoldmanER, 2007. Isolation of anti-toxin single domain antibodies from a semi-synthetic spiny dogfish shark display library. BMC Biotechnol, 7:78.

[5]MiyanabeK, AkibaH, KurodaD, et al., 2018. Intramolecular H-bonds govern the recognition of a flexible peptide by an antibody. J Biochem, 164(1):65-76.

[6]MuraseT, EugenioL, SchorrM, et al., 2014. Structural basis for antibody recognition in the receptor-binding domains of toxins A and B from Clostridium difficile. J Biol Chem, 289(4):2331-2343.

[7]NouriA, AhariH, ShahbazzadehD, 2018. Designing a direct ELISA kit for the detection of Staphylococcus aureus enterotoxin A in raw milk samples. Int J Biol Macromol, 107:1732-1737.

[8]O'KennedyR, 2019. Rapid Antibody-based Technologies in Food Analysis. The United Kingdom by CPI Group (UK) Ltd., Croydon, UK.

[9]OrtegaE, AbriouelH, LucasR, et al., 2010. Multiple roles of Staphylococcus aureus enterotoxins: pathogenicity, superantigenic activity, and correlation to antibiotic resistance. Toxins (Basel), 2(8):2117-2131.

[10]PexaraA, BourrielA, GovarisA, 2010. Staphylococcus aureus and staphylococcal enterotoxins in foodborne diseases. J Hellenic Vet Med Soc, 61(4):316-322.

[11]PinchukIV, BeswickEJ, ReyesVE, 2010. Staphylococcal enterotoxins. Toxins (Basel), 2(8):2177-2197.

[12]PolonelliL, PontonJ, ElguezabalN, et al., 2008. Antibody complementarity-determining regions (CDRs) can display differential antimicrobial, antiviral and antitumor activities. PLoS ONE, 3(6):e2371.

[13]ThomasR, PatenaudeSI, MacKenzieCR, et al., 2002. Structure of an anti-blood group A Fv and improvement of its binding affinity without loss of specificity. J Biol Chem, 277(3):2059-2064.