The Impact of Different B Cell Epitopes on PCV2 Cap's Immunogenicity after Conjugated at C-terminal of Cap

doi:10.7668/hbnxb.20190976

Abstract

Abstract: To explore inexpensive and effective porcine circovirus type 2(PCV2) vaccine, the optimized cap gene of PCV2b(GenBank:AY969004.1) was synthesized according to codon usage of E.coli. Two B cell epitopes of PCV2 (²²⁶LKDPPLNP²³³和¹⁹⁵HVGLGTAF²⁰²) were individually and/or tandemly conjugated at the C-terminal of Cap by PCR technology, then the final segments were cloned into pE-SUMO vector, 3 recombinant vectors were induced after transformed into expression strain BL21 (DE3), identification by SDS-PAGE and Western Blotting, all three recombinant proteins are expressed in soluble form and correctly. The three recombinant proteins were purified by nickel column affinity chromatography, and then SUMO protease was used to remove the fusion tags to obtain the unlabeled Cap-e1, Cap-e2 and Cap-e12 proteins, which were respectively emulsified with equal volume corresponding Freund's adjuvant, twenty 6-week-old female Balb/c mice were randomly divided into 5 groups, immunized with 3 kinds of recombinant proteins, the vaccine group and the PBS group were set as the control group. An immunological evaluation was performed to compare the impact of different B-cell epitopes linked to the C-terminal on the immunogenicity of Cap, the results showed that, except for the second week, the antibody level of each test group was significantly higher than PBS group, and the antibody level produced by the Cap-e12 protein conjugated with 2 epitopes was higher than that of the protein linked only 1 epitope (Cap-e1, Cap-e2), in the sixth week, the antibody level of Cap-e12 protein had no significant difference with the vaccine group. In conclusion, the immunogenicity of Cap is related to the number of linked epitopes, the conjugation of two epitopes can significantly improve the immunogenicity of Cap.

Key words: PCV2, SUMO, Capsid protein, B cell epitope, Immunogenicity

CLC Number:

S852.65

LIU Qingkun, FENG Hua, REN Chunxiao, WEI Qiang, LIU Yunchao, ZHANG Gaiping. The Impact of Different B Cell Epitopes on PCV2 Cap's Immunogenicity after Conjugated at C-terminal of Cap[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(3): 233-238. doi: 10.7668/hbnxb.20190976.

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