Application of Namalva Interferon-α Monoclonal Antibodies for Purification and Enzyme Immnoassay of Interferon-α

Abstract

Namalva (or Namalwa) interferon (IFN)-α was partially purified using a combination of conventional methods and modified acid-ethanol extraction. Four mouse monoclonal antibodies against Namalva IFN-α were prepared by hybridoma technology after immunization with Namalva IFN-α thus purified. Three of these monoclonal antibodies recognized the same or a similar epitope on Namalva IFN-α. One of these antibodies was paired with the fourth recognizing a different epitope and used respectively as enzyme-conjugated antibody and solid-phase antibody in our one step enzyme immunoassay (EIA) for IFN-α. This assay is simple and was able to detect as little as 5 pg of IFN-α in 100 μl of sample in the short time of 5 hr. There was a good correlation between the EIA and bioassay. The use of one of the monoclonal antibodies as an immunoadsorbant to purify Namalva IFN-α is also described.