Induction of immunity in swine by purified recombinant VP1 of foot-and-mouth disease virus.
VP1, a capsid protein of foot-and-mouth disease virus (FMDV), contains neutralizing epitopes of the virus. Due to its poor water solubility, recombinant Escherichia coli derived VP1 (rVP1) has previously been used mainly in a denatured form and is not well characterized. Here, using SDS to assist protein refolding and then removing SDS with a detergent removing column, we have successfully purified rVP1 in two aqueous-soluble forms, i.e. monomer and dimer. Studies showed that dimerization occurs by an inter-molecular disulfide bond between two cysteine residues at position 187 of each monomer. Heat treatment revealed that rVP1 dimer exhibited a more thermal-stable conformation than the monomeric form. Both monomeric and dimeric rVP1 reacted with anti-FMDV antibodies. Immunization studies demonstrated that vaccination of swine with either forms of rVP1 was effective in generating immune responses and protecting them from viral challenge.
Wang
Jeng-Hwan
JH
Institute of Bioagricultural Sciences, Academia Sinica, No. 128 Academia Road, Section 2 Nankang, Taipei 11529, Taiwan.
Liang
Chi-Ming
CM
Peng
Jei-Ming
JM
Shieh
Jeng-Jer
JJ
Jong
Ming-Hwa
MH
Lin
Yeou-Liang
YL
Sieber
Martin
M
Liang
Shu-Mei
SM
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000235:genetics; D055592:Biophysical Phenomena; D001703:Biophysics; D002455:Cell Division / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005536:Foot-and-Mouth Disease / Q000517:prevention & control; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D006358:Hot Temperature; D007114:Immunization; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017510:Protein Folding; D013552:Swine / Q000276:immunology*; D013553:Swine Diseases / Q000517:prevention & control; D013601:T-Lymphocytes / Q000276:immunology; D019444:Vaccines, DNA / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014765:Viral Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic; D014765:Viral Vaccines