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

Holman, Stephen W, McLean, Lynn and Eyers, Claire E ORCID: 0000-0002-3223-5926 (2016) RePLiCal: A QconCAT Protein for Retention Time Standardization in Proteomics Studies. JOURNAL OF PROTEOME RESEARCH, 15 (3). pp. 1090-1102.

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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 to maximize peptide and protein identification rates, and to recalibrate the m/z scale of mass spectrometry data post-acquisition.

Item Type:	Article
Uncontrolled Keywords:	retention time, LC, standardization, SRM, proteomics, QconCAT, mass spectrometry, calibration, quality control
Depositing User:	Symplectic Admin
Date Deposited:	01 Jun 2016 10:00
Last Modified:	19 Jan 2023 07:36
DOI:	10.1021/acs.jproteome.5b00988
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3001434

Available Versions of this Item

• RePLiCal: A QconCAT Protein for Retention Time Standardization in Proteomics Studies. (deposited 31 May 2016 10:31)
  • RePLiCal: A QconCAT Protein for Retention Time Standardization in Proteomics Studies. (deposited 01 Jun 2016 10:00) [Currently Displayed]