Capillary electrophoresis‐mass spectrometry of peptides from enzymatic protein hydrolysis: Simulation and optimization

Abstract

Two important limitations still exist for the characterization of protein digests by capillary electrophoresis-mass spectrometry (CE-MS): (i) the buffer choice (i.e., the buffer must provide an adequate CE separation without ruining the MS signal), and (ii) the frequent generation of "unexpected" peptidic fragments during the enzymatic protein hydrolysis. In this work, a new approach is used to solve these difficulties, namely a theoretical model that relates the electrophoretic behavior of peptides to their sequence. The effectiveness of this procedure is demonstrated by the fast attainment of good CE-MS conditions for analyzing the peptides obtained from an enzymatic protein hydrolysate in a single run. This strategy can provide useful information for helping to characterize "unexpected" fragments from protein digests.

Recent advances in the application of capillary electromigration methods for food analysis

“…the use of CE to analyze proteins from milk and dairy products [33][34][35][36][37][38][39][40][41], the second one would cover the analysis of proteins from cereals (mainly wheat proteins) [42][43][44][45][46][47][48][49], and the third one would include all the other applications [49][50][51][52][53][54][55].…”

“…This includes the detection and quantitation of a bioactive peptide in Vicia narbonensis L. seeds [51], the characterization of proteins from Spirulina platensis microalga using CE-ion trap-mass spectrometry and CE-time of flight-mass spectrometry [52], the CE-MS analysis of basic proteins in foods using a new physically adsorbed polymer coating [53] and the investigation of peptidic maps from proteins using CE-MS and a peptide modelling [54].…”

Recent advances in the application of capillary electromigration methods for food analysis

“…Two novel procedures aimed at determining the amino acid sequence of peptides of interest for the food industry have recently been published. Thus, Simó et al [41] combined the use of a theoretical model that correlates electrophoretic behavior of peptides to their sequence together with CE-MS [44,45] for the study of the cleavage activity of a recombinant pepsin versus its corresponding natural variety using cytochrome C (CytC) as a substrate. To do this, hydrolysates were first analyzed by CE coupled to electrospray ionization mass spectrometry (ESI-MS).…”

Capillary electrophoresis‐electrospray‐mass spectrometry in peptide analysis and peptidomics

“…In this regard, an important limitation arises from the frequent generation of unexpected peptidic fragments during the enzymatic digestion of any protein. To solve this problem, the use of theoretical models to simulate peptide behavior in CE can be very useful [47,48,66,70]. Our group has developed a theoretical model to effectively relate the peptide sequence to their electrophoretic behavior [47].…”