engleski

Accurate identification of microorganisms by de novo sequencing

Majority of currently used LC-MS/MS techniques for protein identification is based on database matching of non-derivatized peptide signals recorded in positive ion mode. Although widely used, such an approach does not always provide satisfactory sequence coverage to unambiguously identify a protein sample. Protein de novo sequencing concept using CAF-/CAF+ reagent (chemically activated fragmentation negative/chemically activated fragmentation positive) enables fast, highly accurate, reliable and easy to use identification of microorganisms down to the species and subspecies level. CAF-/CAF+ (5-formylbenzene-1, 3-disulfonic acid*) is a chemical reagent for the derivatization of peptide samples prior to analysis by tandem mass spectrometry (MS/MS) that enables gathering of both positive and negative peptide ions datasets. Peptide sequences are read from both positive and negative MS/MS spectra and matched against the NCBInr database by developed software named ProteinReader. We present results that show identification of microorganisms down to the subspecies level using either MALDI- or ESI-TOF technique.

Mass spectrometry ; microorganism ; de novo sequencing

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