Your browser doesn't support javascript.
loading
Toward an Ultimate Solution for Peptide Retention Time Prediction: The Effect of Column Temperature on Separation Selectivity.
Villacrés, Carina; Mizero, Benilde; Spicer, Victor; Viner, Rosa; Saba, Julian; Patel, Bhavinkumar; Snovida, Sergei; Jensen, Penny; Huhmer, Andreas; Krokhin, Oleg V.
Afiliação
  • Villacrés C; Manitoba Centre for Proteomics and Systems Biology, Winnipeg R3E 3P4, Canada.
  • Mizero B; Department of Chemistry, University of Manitoba, Winnipeg R3T 2N2, Canada.
  • Spicer V; Manitoba Centre for Proteomics and Systems Biology, Winnipeg R3E 3P4, Canada.
  • Viner R; Thermo Fisher Scientific, San Jose, California 95134, United States.
  • Saba J; Thermo Fisher Scientific, San Jose, California 95134, United States.
  • Patel B; Thermo Fisher Scientific, Rockford, Illinois 61101, United States.
  • Snovida S; Thermo Fisher Scientific, Rockford, Illinois 61101, United States.
  • Jensen P; Thermo Fisher Scientific, Rockford, Illinois 61101, United States.
  • Huhmer A; Thermo Fisher Scientific, San Jose, California 95134, United States.
  • Krokhin OV; Manitoba Centre for Proteomics and Systems Biology, Winnipeg R3E 3P4, Canada.
J Proteome Res ; 23(4): 1488-1494, 2024 04 05.
Article em En | MEDLINE | ID: mdl-38530092
ABSTRACT
We studied the effect of the column temperature on the selectivity of reversed-phase peptide separation in bottom-up proteomics. The number of peptide identifications from 2 h liquid chromatography with tandem mass spectrometry (LC-MS/MS) acquisitions reaches a plateau at 45-55 °C, driven simultaneously by improved separation efficiency, a gradual decrease in peptide retention, and possible on-column degradation of peptides at elevated temperatures. Performing 2D LC-MS/MS acquisitions at 25, 35, 45, and 55 °C resulted in the identification of ∼100,000 and ∼120,000 unique peptides for nonmodified and tandem mass tags (TMT)-labeled samples, respectively. These peptide collections were used to investigate the temperature-driven retention features. The latter is governed by the specific temperature response of individual residues, peptide hydrophobicity and length, and amphipathic helicity. On average, peptide retention decreased by 0.56 and 0.5% acetonitrile for each 10 °C increase for label-free and TMT-labeled peptides, respectively. This generally linear response of retention shifts allowed the extrapolation of predictive models beyond the studied temperature range. Thus, (trap) column cooling from room temperature to 0 °C will allow the retention of an additional 3% of detectable tryptic peptides. Meanwhile, the application of 90 °C would result in the loss of ∼20% of tryptic peptides that were amenable to MS/MS-based identification.
Assuntos
Palavras-chave

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Peptídeos / Espectrometria de Massas em Tandem Idioma: En Revista: J Proteome Res Assunto da revista: BIOQUIMICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Canadá

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Peptídeos / Espectrometria de Massas em Tandem Idioma: En Revista: J Proteome Res Assunto da revista: BIOQUIMICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Canadá