A simple competitive enzyme-linked immunosorbent assay using antigen-beta-galactosidase fusions.
The fusion of the N-terminal 461 bp of the human interferon-alpha 2 (INF) in frame to the beta-galactosidase gene from Escherichia coli is described. The presence of the expected DNA sequence was shown by restriction mapping and DNA sequencing. A fusion protein was demonstrated in crude extracts of E. coli by Western blots using polyclonal anti-beta-galactosidase and monoclonal anti-IFN antibodies. Using monoclonal antibodies specific for the N-terminal region of IFN-alpha and cell-free extracts from an E. coli strain containing the fusion protein, we set up a simple competitive enzyme-linked immunosorbent assay for human interferon. The test described here was linear down to a lower detection limit of at least 1000 Units, or 5 ng human IFN.
D000911:Antibodies, Monoclonal; D000941:Antigens / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005696:Galactosidases / Q000235:genetics / Q000276:immunology; D006801:Humans; D007370:Interferon Type I / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology*; D001616:beta-Galactosidase / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D007370:Interferon Type I; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D005696:Galactosidases; D001616:beta-Galactosidase