ADMET and DMPK  

Peptide retention time prediction for immobilized artificial membrane phosphatidylcholine stationary phase: method development and preliminary observations

Daniel Gussakovsky, Haley Neustaeter, Victor Spicer, Oleg Krokhine

Abstract


Development of the first peptide retention prediction model for immobilized artificial membrane phosphatidylcholine (IAM.PC) stationary phase is reported. 2D LC-MS/MS analysis of a whole cell lysate of S. cerevisiae yielded a retention dataset of ~29,500 tryptic peptides; sufficient for confident assignment of retention coefficients which determine the contribution of individual amino acids in peptide retention. Retention data from the first dimension was used for the modeling: IAM.PC DD2 column, pH 7.4 ammonium bicarbonate, and water/acetonitrile gradient. Peptide separation using IAM.PC was compared to a standard C18 phase (Luna C18(2)). There was a significant reduction in peptide retention (~14% acetonitrile on average), indicating that the phosphatidylcholine stationary phase is significantly more hydrophilic. In comparison to the C18 phase, we found a substantial increase in the relative retention contribution for the positively charged Arg and Lys, and the aromatic Tyr, Trp and His residues. We also observed a decrease in retention contribution for the negatively charged Asp and Glu. This indicates an involvement of electrostatic interactions with the glycerophosphate functional groups, and possibly, delocalization effects from a hydrogen bond between the phosphate group and aromatic side chains in the separation mechanism.

Keywords


Peptide retention modelling; immobilized artificial membrane chromatography

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DOI: http://dx.doi.org/10.5599/admet.520

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ISSN 1848-7718