Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Construction and evaluation of a novel triple cell epitope-based polypeptide vaccine against cow mastitis induced by Staphylococcus aureus, Escherichia coli and Streptococcus

Xiang Liu , Chunlin Chen, Chen Chen, Gregory Marslin, Rui Ding, Sanqiao Wu

Chinese-German Joint Institute for Natural Product Research, College of Biological Science and Engineering, Shaanxi University of Technology; Shaanxi Engineering Research Center of Tall Gastrodia Tuber and Medical Dogwood, Hanzhong 723001, China;

For correspondence:- Xiang Liu Email: liuxiang888525@163.com Tel:+869162641661

Accepted: 19 September 2017 Published: 31 October 2017

Citation: Liu X, Chen C, Chen C, Marslin G, Ding R, Wu S. Construction and evaluation of a novel triple cell epitope-based polypeptide vaccine against cow mastitis induced by Staphylococcus aureus, Escherichia coli and Streptococcus. Trop J Pharm Res 2017; 16(10):2477-2486 doi: 10.4314/tjpr.v16i10.23

© 2017 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To construct a novel triple cell epitope-based polypeptide vaccine against cow mastitis induced by Staphylococcus aureus, Escherichia coli and Streptococcus and to reduce the use of antibiotics.

Methods: Based on bioinformatics approach, a novel triple epitope-based polypeptide (CM-TEP) was designed and subjected to Ni-NTA flow resin purification. Purified CM-TEP was immunized into mice to prepare a polyclonal antibody. Pull-down assays and enzyme-linked immunosorbent assay (ELISA) were used to detect the interaction between CM-TEP antibodies and S. aureus, E. coli and Streptococcus. Active immunity mice and challenge of bacterial pathogens were used to detect immune protection of CM-TEP. Additionally, the optimal expressing conditions of CM-TEP strain were analyzed using orthogonal test design.

Results: A novel cow mastitis triple cell epitope-based polypeptide (CM-TEP) with a MW of 36 kDa was designed, purified and used to immunize mice to prepare a polyclonal antibody. Pull-down assays and ELISA data showed that CM-TEP antibodies directly interacted with S. aureus, E. coli and Streptococcus. CM-TEP displayed a significant immune protective effect against infection by S. aureus (50 %, p < 0.05) and E. coli (54.54 %, p < 0.05) and provided some immune protective effect (30.78 %, p > 0.05) against Streptococcus. The optimum expressing conditions of CM-TEP were as follows: IPTG concentration of 0.3 mmol/L, strain OD600 value of 1, inducing temperature of 37 ^oC, and inducing time of 8 h.

Conclusion: The findings suggest that epitope-based vaccine of CM-TEP may be a useful strategy for treating cow mastitis induced by S. aureus, E. coli and Streptococcus

Keywords: Cow mastitis, Epitope vaccine, Immunogenicity, Immune protective