A method of detecting the activity of toxins produced by the Peptostreptococcaceae and Clostridiaceae families using mass spectrometry with desorption ionization

Abstract

The method of detecting the activity of the toxin produced by the bacteria of Peptostreptococcaceae or Clostridiaceae family using mass spectrometry, which includes the following steps: a) providing a planar surface suitable for measuring using mass spectrometry with bound affinity molecules; b) incubating the planar surface from step a) with the substrate of toxin to bound the substrate to affinity molecules, where the substrate is a recombinant marked RhoA protein, which is prepared using expression plasmid p-GEX-6P1; c) the subsequent incubation of the planar surface from step b) with the sample at the presence of modifying compound to form modified substrate, possibly unmodified substrate, if the toxin in the sample is not active; d) removing the residues of the sample; e) subjecting the modified / unmodified substrate on the planar surface to the matrix-assisted laser desorption / ionising, its ionising and subsequent measuring of mass spectrum of the modified / unmodified substrate; f) analysing the measured mass spectrum to detect the active toxin in the sample.