Analysis of intact venom proteins with capillary zone electrophoresis - mass spectrometry

• Published in: Microchemical journal 2024-05, Vol.200, p.110290, Article 110290
• Main Authors: Vishwakarma, Gayatri, Andrasi, Melinda, Szabo, Ruben, Hajdu, Peter, Petrilla, Vladimir, Petrillová, Monika, Legath, Jaroslav, Gaspar, Attila
• Format: Article
• Language: English
• Subjects: Capillary zone electrophoresis; Intact protein; Mass spectrometry; Snake venom; Top down proteomics
• ISSN: 0026-265X
• DOI: 10.1016/j.microc.2024.110290

[Display omitted] •Venom provides a characteristic feature (fingerprint) of a given snake.•CZE-MS was applied for the intact protein analysis of similar venom samples.•Using 1 M formic acid as BGE minimal adsorption and narrow peaks were obtained.•More than 250 neuropeptides were detected in the venoms of different subspecies.•Venoms of the same subspecies gained from different locations include similar proteins.•Venoms collected from different subspecies exhibit different protein patterns. In this work we demonstrated the favourable analytical performance of capillary zone electrophoresis coupled with mass spectrometry (CZE-MS) for the intact protein analysis of similar venom samples. Venom provides a characteristic feature (fingerprint) of a given snake. Using 1 M formic acid (pH = 1.9) as background electrolyte, minimal adsorption and narrow peaks shapes - thus good separation efficiencies - were obtained for the protein components of the venom samples. The precision of migration times and peak areas were 1.9–2.8 RSD% and 0.8–7.2 RSD%, respectively and the theoretical plate numbers were 32000–238000 for peaks having signal-to-noise ratio (S/N) larger than 50. More than 250 different neuropeptides (7–10 kDa) were detected in the venoms obtained from snakes of 9 different subspecies (belonging either to Naja or Dendroaspis species). The protein contents of the venoms of the same subspecies collected from different geographical regions are similar and differ only in a few (less than 10 %) components. However, the venoms collected from different subspecies (but from the same species) exhibit very different protein patterns: no matches were found in the molecular weights of proteins.