An enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies to different parts of the gram-negative lipopolysaccharide core region |
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Authors: | B J Appelmelk A M Verweij-Van Vught D M MacLaren L G Thijs |
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Institution: | 1. Research Group for Commensal Infections, Vrije Universiteit, School of Medicine, Amsterdam, The Netherlands;2. Department of Acute Internal Medicine, Virje Universiteit, School of Medicine, Amsterdam, The Netherlands |
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Abstract: | Bovine serum albumin was complexed with the core antigens of either Escherichia coli J5 LPS, Salmonella minnesota R595 LPS or E. coli lipid A. These core-BSA complexes were used for solid-phase coating in ELISAs for anti-core antibodies. Antibodies, binding to various parts of the core region were easily quantified in a single experimental set-up, which was hitherto not possible. The ELISA has only 3 incubation steps and is not costly as only moderate amounts of the core antigens (i.e., 1 microgram per test) were needed for coating. The sensitivity proved to be excellent and the complexes were biologically fully active (compared to native, smooth LPS), which make them suitable for the screening (after fusion) of monoclonal anti-core antibodies. Another possible application is the large-scale screening of blood-bank sera in order to find samples with a high anti-core antibody content. |
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Keywords: | enzyme-linked immunosorbent assay core antigen-bovine serum albumin complexes polyclonal and monoclonal antisera Gram-negative core region ELISA enzyme-linked immunosorbent assay BSA bovine serum albumin LPS lipopolysaccharide PBS phosphate-buffered saline CFU colony-forming unit SD standard deviation TEA triethylamine PH passive hemolysis PHA passive hemagglutination OD optical density |
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