PXD058240

PXD058240 is an original dataset announced via ProteomeXchange.

Title	Novel activity assay for botulotoxin A1 detection using functionalized chips and matrix-assisted laser desorption/ionization mass spectrometry
Description	Botulinum neurotoxins (BoNTs) are a group of neurotoxins produced by Clostridium bacteria. Their effect on neuro-muscular connections through cleaving proteins of the SNARE complex results in blocking acetylcholine signal transduction. To prevent external intoxication by BoNT, analysis of potential sources of BoNT needs to be performed. Currently available methods target the presence of BoNTs molecules but only a fraction is screening real toxic activity. The FDA-approved mouse bioassay, based on exposing live mice to the potentially contaminated food, is the most common. Unfortunately, this assay is expensive, time consuming and ethically questionable. That is why there is a need for assays that can measure the activity of BoNTs enzymatically. We present an assay combining known EndoPep-MS assay with detection protein affinity chips manufactured by ion soft-landing technology. The toxic activity is indirectly determined by monitoring their endopeptidase activity as a change in molecular mass of cleaved substrate peptides exposed to BoNTs. Our modification utilizes a protein array modified by affinity molecules towards either the BoNTs or the substrate peptides. Both variants allow to perform the in-situ reaction and detection of substrate peptides by MALDI-ToF MS on the protein chip. This method demonstrated successful detection of active BoNT/A1 in both buffer and complex matrices, achieving a detection limit of 0.5 ng/mL.
HostingRepository	PRIDE
AnnounceDate	2025-03-21
AnnouncementXML	Submission_2025-03-21_01:59:21.767.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zuzana Kalaninova
SpeciesList	scientific name: Clostridium botulinum; NCBI TaxID: 1491;
ModificationList	No PTMs are included in the dataset
Instrument	autoflex

Revision	Datetime	Status	ChangeLog Entry
0	2024-11-25 07:35:05	ID requested
⏵ 1	2025-03-21 01:59:22	announced

10.1080/14789450.2025.2482933;

submitter keyword: neurotoxin, MALDI-ToF, activity,botulinum, EndoPep, assay

Petr Pompach
contact affiliation	Institute of Biotechnology, The Czech Academy of Sciences, Vestec, Czech Republic
contact email	petr.pompach@ibt.cas.cz
lab head
Zuzana Kalaninova
contact affiliation	Faculty of Science, Charles University
contact email	kalaninz@natur.cuni.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD058240
     1. Label: PRIDE project
     2. Name: Novel activity assay for botulotoxin A1 detection using functionalized chips and matrix-assisted laser desorption/ionization mass spectrometry