Production of egg yolk antibody (IgY) against recombinant canine parvovirus VP2 protein
Han, ShuizhongZhang, XiaoyingZhao, Jinzi
Background: Canine parvovirus-2 (CPV-2) disease is highly contagious and often fatal in canine with symptoms of hemorrhagic diarrhea and myocarditis. Three variants, CPV-2a, CPV-2b and CPV-2c, had emerged and replaced the CPV-2 in the past decades worldwide. Emergences of the new variants may increase the difficulty of CPV diagnosis and treatment. CPV capsid consists of 60 copies of a combination of viral coat proteins VP1 (82 kDa), VP2 (67 kDa) and VP3 (63.5 kDa), while about 90% of the capsid proteins are VP2 and most of the B cell epitopes are located on the VP2. IgY technology has been recognized as a promising alternative to generate large amount of qualified high specific antibody for use in immunodiagnostic and in immunotherapy with the advantages of relatively simple, noninvasive method and large-scale production over mammalian antibody. Furthermore, IgY antibody does not react with mammalian IgG nor binding to the rheumatoid factor, which may reduce the false positive results in immunoassays. Anti-VP2 IgY antibody has never been reported before, here we describe a method for the production of anti-VP2 IgY, which could be applied in the diagnosis and treatment for the CPV infection.Materials, Methods & Results: A CPV strain was isolated from a clinical sample. The VP2 ORF (Open reading frame, ORF) was amplified by PCR and inserted into the pMD18-T clone vector. The
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