Using distilled water for the extraction of mucosal antibodies and the subsequent application in RSV neutralization test.
Virus neutralization (VN) is an important functional test for evaluating RSV vaccines, also encompassing in mucosal secretion of the respiratory tract considering the infection route. In our previous study, an immunoglobin extraction method described by Bergquist et al. was adopted for RSV ELISA, but it was not suitable for virus neutralization test due to the cell toxicity of the 2% saponin solution used for the antibody extraction. In order to overcome this problem, several solvents including distilled water were tested in the present study for the capacity to extract immunogloblins. Antibodies in the extracts were evaluated and compared by ELISA. Distilled water was as efficient as the 2% saponin solution for extraction of total IgA, RSV specific IgA and IgG. More importantly, the organ extracts obtained subsequently could be used for virus neutralization test without causing adverse effect on the cell culture. Therefore, distilled water was finally chosen as the solvent for immunoglobulin extraction from mucosal organs when both ELISA and virus neutralization test are required.
Swedish University of Agricultural Sciences, College of Veterinary Medicine, Department of Veterinary Microbiology, Uppsala, Sweden.
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D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016882:ISCOMs / Q000008:administration & dosage / Q000276:immunology; D007070:Immunoglobulin A / Q000302:isolation & purification*; D007074:Immunoglobulin G / Q000302:isolation & purification; D008168:Lung / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000276:immunology; D009297:Nasal Mucosa / Q000276:immunology*; D009500:Neutralization Tests / Q000379:methods*; D012136:Respiratory Syncytial Viruses / Q000276:immunology*; D012503:Saponins / Q000494:pharmacology; D018709:Statistics, Nonparametric; D014867:Water
D000914:Antibodies, Viral; D016882:ISCOMs; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D012503:Saponins; D014867:Water