Preparation And Characterization Of Porcine A-interferon Monoclonal Antibody

Interferon alpha (IFN-α) is a glycoprotein with high bioactivity, which is produced by induced cells. It possesses Broad-spectrum anti-viral activity for the same kinds of animal cells. At present, it has been confirmed that nearly a hundred of diseases are infected to human through animal, most of which are viral infectious diseases. However, effective vaccines and drugs are not available for prevention and treatment of the diseases. Furthermore, biological characteristics of some pathogenic virus are still unclear. Among all IFN family member, IFN-αhas the most anti-viral capacity, but its application in medical science and veterinary science is currently limited. Thence IFN-αresearch embodies important practical significance. Anti-IFN-αmonoclonal antibody development can lay the foundation for evaluation of animal host anti-virus level and molecular mechanism studies.In this study, use hybridoma technology, the immuned spleen cells prepared from BALB / c mice vaccinated recombinant protein of pIFN-αunder sterile conditions, and fused with myeloma cell of SP2/0, through a series cloning of culture and di-system screening of indirect ELISA, three hybridoma cell lines secreting stabily anti-IFN-αmonoclonal antibody(McAb) have been obtained successfully. After subtype identification of McAb, displayed one is IgG2a, two are IgG2b. The chromosome of hybridoma cell lines secreting McAb was counted and found that the average is 84, and chromosomes of the myeloma cell line only is the average of 50, this demonstrate that hybridoma cell lines obtained on the genetic point; By ELISA detection McAb titer of hybridoma, three hybridoma lines all can reach 1:105 in ascites, and is 1:104 in the cell supernatant; Monoclonal antibody antigen-binding site analysis showed that three of antigen antibody binding sites are basically the same; SDS-PAGE analysis of McAb shows heavy chain molecules of three lines are more than 60ku, light chain molecules are more than 30ku, and obtained higher purity McAb; The stability testing of the hybridoma can draw conclusion that the three cells antibody titer is between 1:104 ~ 1:105 after 4 times cloning culture, it shows that three hybridoma cells all possess a good genetic stability.