Novel application of Influenza A virus-inoculated chorioallantoic membrane to characterize a NP-specific monoclonal antibody for immunohistochemistry assaying
- Author:
Yang Chang TU
1
;
Kuang Yu CHEN
;
Chung Kung CHEN
;
Ming Chu CHENG
;
Shu Hwae LEE
;
Ivan Chen CHENG
Author Information
- Publication Type:Original Article
- Keywords: Chorioallantoic membrane; Immunohistochemistry; Influenza A virus; Antibodies, Monoclonal; Nucleoproteins
- MeSH: Animals; Antibodies, Monoclonal; Blotting, Western; Chickens; Chorioallantoic Membrane; Diagnosis; Disease Outbreaks; Epitopes; Fluorescent Antibody Technique; Formaldehyde; Immunohistochemistry; Influenza A virus; Influenza in Birds; Influenza, Human; Nucleoproteins; Taiwan
- From:Journal of Veterinary Science 2019;20(1):51-57
- CountryRepublic of Korea
- Language:English
- Abstract: Monoclonal antibodies (MAbs) are widely applied in disease diagnoses. Herein, we report a MAb, WF-4, against Influenza A virus nucleoprotein (NP), its broad response with Influenza A virus, and its application in an immunohistochemistry (IHC) assay. WF-4 was screened by immunofluorescence assay (IFA). The results showed that its reactivity with baculovirus-expressed full-length recombinant NP (rNP) in Western blot (WB), indicating its IHC applicability. Fifteen Influenza A virus (reference subtypes H1 to H15) infected chicken embryonated chorioallantoic membranes (CAM), fixed by formalin, were all detectable in the WF-4-based IHC assay. Also, the reactivity of the IHC test with NP from experimentally inoculated H6N1 and from all recent outbreaks of H5 subtype avian Influenza A virus (AIV) field cases in Taiwan showed positive results. Our data indicate that CAM, a by-product of Influenza A virus preparation, is helpful for Influenza A virus-specific MAb characterization, and that the WF-4 MAb recognizes conserved and linear epitopes of Influenza A virus NP. Therefore, WF-4 is capable of detecting NP antigens via IHC and may be suitable for developing various tests for diagnosis of Influenza A virus and, especially, AIV infection.
