Karine Bagramyan , Ph.D., is a staff scientist in the laboratory of Professor Markus Kalkum at the Department of Molecular Imaging and Therapy of Diabetes and Metabolism Research Institute of City of Hope in Duarte, CA. Her research involves the development of mass spectrometry-based targeted proteomics methods, and their application for botulinum neurotoxin detection, evaluation, and quantitation. She began using Skyline in 2019 to analyze LC-MS/MS (MRM and PRM) data dealing with a large number of samples for targeted quantitative analysis.
Using Skyline to Quantify Botulinum Neurotoxin Activity in Complex Biological Samples
Botulinum neurotoxins (BoNTs) are the most potent toxins known, with the lethal dose for mice (MLD50) of ~33 amol. Due to BoNT’s extraordinary toxicity, BoNT detection assays have to be highly sensitive and capable of detecting toxin concentrations equal or below one MLD50 in biological matrices. We have applied high-resolution PRM and MRM LC/MS techniques to quantify BoNTs in human serum. Our methodology is based on the detection of BoNT’s proteolytic activity and does not require BoNT-specific antibodies. This presentation will highlight Skyline’s utility for the design and optimization of our PRM and MRM assays.
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This brilliant software provided us with a solid bioinformatics pipeline for the entire project: From the generation of calibration curves using stable isotope-labeled synthetic peptide standards, to the quantification of attomolar concentrations of BoNT, resulting in a novel assay that has unmatched limits of detection and quantification.
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