Preparation And Characterization Of Monoclonal Antibody Against Interferon-α Of Goose

People focused to research molecular composition, gene expression and bioactivities of interferon(IFN), after Isaacs and Lindenmann from England found IFN in 1957.IFN that play a role in antitumor and antivirus was low molecular weight dissoluble sugar protein.IFN includesⅠIFN(including IFN-αand IFN-β)andⅡIFN(i.e. IFN-γ). IFN-αplayed an important role in antivirus. IFN-αis from leucocyte, Namalwa cell, KG-1 cell, B lymphocyte, chronic myelogenous leukemia cell and fibroblast induced by newcastle disease virus cell and vesicular stomatitis virus.Balb/C mice were immunized with the recombinant plasmid pcDNA3.1-GoIFN-α. The screening McAbs against interferon-αof goose was performed with recombinant bacterium pET-30а-GoIFN-α/BL21. SP2/0 was fused with spleen cells from Balb/C mice immunized. The hybridoma supernatants were delected by indirect ELISA. The positive cells were subcloned 3~5 times by limiting dilution. Two strains hybridoma cells line steadily secreting McAbs against interferon-αof goose were obtained and named 2H6, 4C8 respectively.The chromosomal number of hybridoma cell were 103 and 89 respectively. The subtype of both 2 McAbs were IgM and the light chains areκchain;and the ELISA titers of two McAbs ascitic fluids were up to 10-4. Western blot analysis showed that the two McAbs could react with recombinant IFN-α, and one of McAbs could react with natual interferon. We estimated that two McAbs may be aimed at different epitopes. The hybridoma cell strains grew well after continuous cultivation or after being lyophilized , and the titers of secreted McAbs were stable. McAb neutrality activity showed that McAb 4C8 could neutralized goose natual interferon. Neutrality activity of McAb showed that McAb 4C8 could neutralized anti-virus action of narural interferon.In a word, two McAbs were specific to the goose interferonαare obtained, which provided an important method to study and examine goose interferonα; and have application value in further studies on immune detection, the functions of immune cells and immune regulation.