Development of a double-recombinant antibody sandwich ELISA for quantitative detection of epsilon toxoid concentration in inactivated Clostridium perfringens vaccines

BMC Veterinary Research

Mehdi Golchin¹  Maryam Alibeiki²  Mohammad Tabatabaei² 

[1] Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran;Department of Pathobiology, Faculty of Veterinary Medicine, Shiraz University, Shiraz, Iran;

关键词: Epsilon toxin;    Clostridium perfringens;    Recombinant antibody;    Phage display;    Sandwich ELISA;   

DOI  :  10.1186/s12917-020-02572-4

来源: Springer

【 摘 要 】

BackgroundEpsilon toxin (ETX) causes a commonly fatal enterotoxemia in domestic animals. Also, ETX causes serious economic losses to animal husbandry. In this study, we selected several clones against ETX using repertoires displayed on filamentous phage. Anti-ETX specific clones were enriched by binding to immobilized antigen, followed by elution and re-propagation of phage. After multiple rounds of binding selection, ELISA analysis showed that most isolated clones had high affinity and specificity for ETX.ResultsTwo recombinant monoclonal antibodies against ETX were isolated by phage display technology. B1 phage VH antibody isolated from DAb library and G2 soluble scFv antibody isolated from Tomlinson I + J libraries have been applied as the capture and detection antibodies for developing an ETX sandwich ELISA test, respectively.ConclusionsDesigned ETX sandwich ELISA could be a valuable tool for quantitative detection of ETX in inactivated commercial vaccines against enterotoxemia.Graphical abstract

【 授权许可】

CC BY