RePLiCal: A QconCAT Protein for Retention Time Standardization in Proteomics Studies

Journal of Proteome Research
Stephen W HolmanClaire E Eyers

Abstract

This study introduces a new reversed-phase liquid chromatography retention time (RT) standard, RePLiCal (Reversed-phase liquid chromatography calibrant), produced using QconCAT technology. The synthetic protein contains 27 lysine-terminating calibrant peptides, meaning that the same complement of standards can be generated using either Lys-C or trypsin-based digestion protocols. RePLiCal was designed such that each constituent peptide is unique with respect to all eukaryotic proteomes, thereby enabling integration into a wide range of proteomic analyses. RePLiCal has been benchmarked against three commercially available peptide RT standard kits and outperforms all in terms of LC gradient coverage. RePLiCal also provides a higher number of calibrant points for chromatographic retention time standardization and normalization. The standard provides stable RTs over long analysis times and can be readily transferred between different LC gradients and nUHPLC instruments. Moreover, RePLiCal can be used to predict RTs for other peptides in a timely manner. Furthermore, it is shown that RePLiCal can be used effectively to evaluate trapping column performance for nUHPLC instruments using trap-elute configurations, to optimize gradients t...Continue Reading

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Citations

Sep 21, 2016·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Stephen W HolmanRobert J Beynon
Apr 4, 2017·Scientific Reports·Richard J BennettRobert J Beynon
Oct 22, 2017·Molecular & Cellular Proteomics : MCP·Nobuaki TakemoriRobert J Beynon
Aug 2, 2017·Mass Spectrometry Reviews·Francisco Calderón-CelisAlfredo Sanz-Medel
Nov 16, 2019·Proteomics·Constance A SobseyChristoph H Borchers
Aug 13, 2017·Mass Spectrometry Reviews·Wout BittremieuxKris Laukens
Dec 5, 2019·Frontiers in Pharmacology·Adriana Muñoz-TalaveraAna Rosa Rincón-Sánchez
Jan 23, 2019·Journal of Proteome Research·Katie J LeeElizabeth G Canty-Laird
Dec 7, 2018·Journal of Proteome Research·Christopher M RoseDonald S Kirkpatrick

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