A simple competitive enzyme-linked immunosorbent assay using antigen-beta-galactosidase fusions |
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Authors: | A Peterhans M Mecklenburg F Meussdoerffer K Mosbach |
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Institution: | Institute of Biotechnology, Swiss Federal Institute of Technology, Zurich. |
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Abstract: | 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. |
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