Sensitive detection of botulinum neurotoxin types C and D with an immunoaffinity chromatographic column test |
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Authors: | Gessler Frank Hampe Katrin Böhnel Helge |
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Affiliation: | Institute for Applied Biotechnology in the Tropics, Kellnerweg 6, 37077 G?ttingen, Germany. fgessle@gwdg.de |
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Abstract: | A sensitive and specific immunoassay for the simultaneous detection of Clostridium botulinum type C (BoNT/C) and type D neurotoxin was developed. Goat anti-mouse immunoglobulin G was bound to polyethylene disks in a small disposable column used for this assay. The sample was preincubated together with monoclonal antibodies specific for the heavy chain of BoNT/C and D and affinity-purified, biotinylated polyclonal antibodies against these neurotoxins. This complex was captured on the assay disk. Streptavidin-poly-horseradish peroxidase was used as a conjugate, and a precipitating substrate allowed the direct semiquantitative readout of the assay, if necessary. For a more accurate quantitative detection, the substrate can be eluted and measured in a photometer. Depending on the preincubation time, a sensitivity of 1 mouse lethal dose ml(-1) was achieved in culture supernatants. |
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