Heterologous expression of full-length capsid protein of porcine circovirus 2 in Escherichia coli and its potential use for detection of antibodies.
A capsid protein of porcine circovirus 2 (PCV 2) serves as a diagnostic antigen for the detection of PCV 2-associated disease known as a postweaning multisystemic wasting syndrome (PMWS). In this report, a bacterial expression system was developed for the expression and purification of the full-length PCV 2 capsid (Cap) protein from a codon-optimized cap gene. Replacement of rare arginine codons located at the 5' end of the cap reading frame with codons optimal for E. coli was found to overcome the poor expression of the viral protein in the prokaryotic system. The Cap protein was purified to greater than 95% homogeneity by using a single cation-exchange chromatography at a yield of 10 mg per litre of bacterial culture. Despite the failure of the E. coli-expressed Cap protein to self-assemble into virus-like particles (VLPs), the immunization of mice with recombinant Cap yielded antibodies with the same specificity as those raised against native PCV 2 virions. In addition, the antigenic properties of the purified Cap protein were employed in a subunit-based indirect ELISA to monitor the levels of PCV 2 specific antibodies in piglets originating from a herd which was experiencing PCV 2 infection. These results pave the way for a straightforward large-scale production of the recombinant PCV 2 capsid protein and its use as a diagnostic antigen or a PCV 2 subunit vaccine.
Marcekova
Zuzana
Z
Proteix s. r. o., Nad Safinou II/365 Vestec, 252 42 Jesenice u Prahy, Czech Republic.
Psikal
Ivan
I
Kosinova
Eva
E
Benada
Oldrich
O
Sebo
Peter
P
Bumba
Ladislav
L
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017925:Circovirus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007114:Immunization; D008969:Molecular Sequence Data; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000175:diagnosis* / Q000517:prevention & control / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013552:Swine / Q000821:virology*; D013553:Swine Diseases / Q000175:diagnosis* / Q000517:prevention & control / Q000821:virology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology; D014771:Virion / Q000378:metabolism
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; D014765:Viral Vaccines