Production of an Hybridoma Library to Recombinant Porcine Alpha I Interferon: A Very Sensitive Assay (ISBBA) Allows the Detection of a Large Number of Clones

The preparation of a panel of hybridoma cell lines secreting monoclonal antibodies (MAbs) specific for Recombinant Porcine Alpha I Interferon is reported. Of these MAbs, 28 were subcloned and 21 secreted their antibody during several months. They were partially characterized for their ability to bind and neutralize Alpha Interferons (IFNs-α) from different animal species. All the clones bound Recombinant Porcine IFN-α₁, 8 bound Human leukocyte IFN, and one bound recombinant Human IFN-α_2b, but not IFN-α_2a. Two screening procedures were used for the detection of specific MAbs: neutralization assay and immunosorbent binding bioassay. This last method appears to be simple and very sensitive since it permits to detect 100pg/ml MAb. In addition it can detect weakly, as well as strongly neutralizing antibodies. Probable mechanisms involved in this assay, and possible applications of this method for IFNs-α subtyping are discussed.