Sensitive detection of botulinum neurotoxin types C and D with an immunoaffinity chromatographic column test.
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.
Gessler
Frank
F
Institute for Applied Biotechnology in the Tropics, Kellnerweg 6, 37077 Göttingen, Germany. fgessle@gwdg.de
Hampe
Katrin
K
Böhnel
Helge
H
D001905:Botulinum Toxins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D003014:Clostridium botulinum / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007118:Immunoassay; D007700:Kinetics; D008970:Molecular Weight; D012680:Sensitivity and Specificity
C096325:botulinum toxin type D; D001905:Botulinum Toxins; C096324:botulinum toxin type C