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Web of Proceedings - Francis Academic Press
Web of Proceedings - Francis Academic Press

Prokaryotic Expression and Polyclonal Antibody Preparation of Cdtc Protein from Haemophilus Parasuis

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DOI: 10.25236/isbsbh.2020.020


Aihua Chen

Corresponding Author

Aihua Chen


Haemophilus parasuis is the pathogen of swine Glasser’s disease which responsible for enormous economic losses in the swine industry. Cytolethal distending toxin (CDT), containing three subunits: CdtA, CdtB and CdtC, is an important virulence factor of H. parasuis. With the character of decreased cell toxicity by mutation, CdtC is suitable for production of antibodies and it can be successfully expressed in soluble form in Escherichia coli when the signal peptides were removed. In this study, CdtC expression plasmid was constructed and transformed into E. coli BL21 (DE3). CdtC protein was expressed in large quantities and purified by nickel affinity chromatography. We obtained and verified CdtC protein with purity > 85% by SDS-PAGE and Western Blot. Purified CdtC protein then was used for animal immunization. Specific antibodies produced by the experimental animals are collected. Finally, polyclonal antibodies with a titer ≥6.4×104 was obtained and confirmed by ELISA


Haemophilus parasuis, Cdtc, Polyclonal antibodies