LFQuant: A label‐free fast quantitative analysis tool for high‐resolution LC‐MS/MS proteomics data

Database searching based methods for label‐free quantification aim to reconstruct the peptide extracted ion chromatogram based on the identification information, which can limit the search space and thus make the data processing much faster. The random effect of the MS/MS sampling can be remedied by cross‐assignment among different runs. Here, we present a new label‐free fast quantitative analysis tool, LFQuant, for high‐resolution LC‐MS/MS proteomics data based on database searching. It is designed to accept raw data in two common formats (mzXML and Thermo RAW), and database search results from mainstream tools (MASCOT, SEQUEST, and X!Tandem), as input data. LFQuant can handle large‐scale label‐free data with fractionation such as SDS‐PAGE and 2D LC. It is easy to use and provides handy user interfaces for data loading, parameter setting, quantitative analysis, and quantitative data visualization. LFQuant was compared with two common quantification software packages, MaxQuant and IDEAL‐Q, on the replication data set and the UPS1 standard data set. The results show that LFQuant performs better than them in terms of both precision and accuracy, and consumes significantly less processing time. LFQuant is freely available under the GNU General Public License v3.0 at http://sourceforge.net/projects/lfquant/.

[1]  P. Holland,et al.  Robust regression using iteratively reweighted least-squares , 1977 .

[2]  Craig Lawless,et al.  Global absolute quantification of a proteome: Challenges in the deployment of a QconCAT strategy , 2011, Proteomics.

[3]  Jie Ma,et al.  Bayesian Nonparametric Model for the Validation of Peptide Identification in Shotgun Proteomics*S , 2009, Molecular & Cellular Proteomics.

[4]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[5]  J. Koziol,et al.  Label-free, normalized quantification of complex mass spectrometry data for proteomics analysis , 2009, Nature Biotechnology.

[6]  K. Resing,et al.  Comparison of Label-free Methods for Quantifying Human Proteins by Shotgun Proteomics*S , 2005, Molecular & Cellular Proteomics.

[7]  J. Yates,et al.  An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database , 1994, Journal of the American Society for Mass Spectrometry.

[8]  Ouyang Chen-xing Reversible retention time alignment algorithm based on local regression , 2011 .

[9]  Jianqi Li,et al.  A new strategy to filter out false positive identifications of peptides in SEQUEST database search results , 2007, Proteomics.

[10]  Lukas N. Mueller,et al.  SuperHirn – a novel tool for high resolution LC‐MS‐based peptide/protein profiling , 2007, Proteomics.

[11]  M. Mann,et al.  MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification , 2008, Nature Biotechnology.

[12]  Lennart Martens,et al.  A comparison of MS2‐based label‐free quantitative proteomic techniques with regards to accuracy and precision , 2011, Proteomics.

[13]  Guanghui Wang,et al.  Label-free protein quantification using LC-coupled ion trap or FT mass spectrometry: Reproducibility, linearity, and application with complex proteomes. , 2006, Journal of proteome research.

[14]  Michael J MacCoss,et al.  Comparison of database search strategies for high precursor mass accuracy MS/MS data. , 2010, Journal of proteome research.

[15]  B. Vanhaesebroeck,et al.  Quantitative Profile of Five Murine Core Proteomes Using Label-free Functional Proteomics*S , 2007, Molecular & Cellular Proteomics.

[16]  Mehdi Mirzaei,et al.  Less label, more free: Approaches in label‐free quantitative mass spectrometry , 2011, Proteomics.

[17]  Fuchu He,et al.  Relationship between sample loading amount and peptide identification and its effects on quantitative proteomics. , 2009, Analytical chemistry.

[18]  M. Mann,et al.  Exponentially Modified Protein Abundance Index (emPAI) for Estimation of Absolute Protein Amount in Proteomics by the Number of Sequenced Peptides per Protein*S , 2005, Molecular & Cellular Proteomics.

[19]  E. Marcotte,et al.  Absolute protein expression profiling estimates the relative contributions of transcriptional and translational regulation , 2007, Nature Biotechnology.

[20]  Mona Singh,et al.  Protein quantification across hundreds of experimental conditions , 2009, Proceedings of the National Academy of Sciences.

[21]  Olivier Langella,et al.  MassChroQ: A versatile tool for mass spectrometry quantification , 2011, Proteomics.

[22]  Linfeng Wu,et al.  Role of spectral counting in quantitative proteomics , 2010, Expert review of proteomics.

[23]  Birgit Schilling,et al.  Repeatability and reproducibility in proteomic identifications by liquid chromatography-tandem mass spectrometry. , 2010, Journal of proteome research.

[24]  Lewis Y. Geer,et al.  DBParser: web-based software for shotgun proteomic data analyses. , 2004, Journal of proteome research.

[25]  Michael K. Coleman,et al.  Statistical analysis of membrane proteome expression changes in Saccharomyces cerevisiae. , 2006, Journal of proteome research.

[26]  Chih-Chiang Tsou,et al.  IDEAL-Q, an Automated Tool for Label-free Quantitation Analysis Using an Efficient Peptide Alignment Approach and Spectral Data Validation* , 2009, Molecular & Cellular Proteomics.

[27]  R. Aebersold,et al.  Proteomics: the first decade and beyond , 2003, Nature Genetics.

[28]  Pei Wang,et al.  Bioinformatics Original Paper a Suite of Algorithms for the Comprehensive Analysis of Complex Protein Mixtures Using High-resolution Lc-ms , 2022 .

[29]  Natalie I. Tasman,et al.  A guided tour of the Trans‐Proteomic Pipeline , 2010, Proteomics.

[30]  Jie Ma,et al.  Improving the sensitivity of MASCOT search results validation by combining new features with Bayesian nonparametric model , 2010, Proteomics.

[31]  Benjamin Thomas,et al.  Comparative evaluation of label‐free SINQ normalized spectral index quantitation in the central proteomics facilities pipeline , 2011, Proteomics.

[32]  R. Beavis,et al.  A method for reducing the time required to match protein sequences with tandem mass spectra. , 2003, Rapid communications in mass spectrometry : RCM.

[33]  Robertson Craig,et al.  TANDEM: matching proteins with tandem mass spectra. , 2004, Bioinformatics.

[34]  M. Mann,et al.  More than 100,000 detectable peptide species elute in single shotgun proteomics runs but the majority is inaccessible to data-dependent LC-MS/MS. , 2011, Journal of proteome research.

[35]  Jianqiu Zhang,et al.  MRCQuant- an accurate LC-MS relative isotopic quantification algorithm on TOF instruments , 2011, BMC Bioinformatics.

[36]  J. Yergey A GENERAL APPROACH TO CALCULATING ISOTOPIC DISTRIBUTIONS FOR MASS SPECTROMETRY. , 1983, Journal of mass spectrometry : JMS.

[37]  Borjana Arsova,et al.  Precision, Proteome Coverage, and Dynamic Range of Arabidopsis Proteome Profiling Using 15N Metabolic Labeling and Label-free Approaches , 2012, Molecular & Cellular Proteomics.

[38]  Jacob D. Jaffe,et al.  PEPPeR, a Platform for Experimental Proteomic Pattern Recognition*S , 2006, Molecular & Cellular Proteomics.

[39]  Rachel M. Adams,et al.  Systematic comparison of label-free, metabolic labeling, and isobaric chemical labeling for quantitative proteomics on LTQ Orbitrap Velos. , 2012, Journal of proteome research.

[40]  David L. Tabb,et al.  Performance Metrics for Liquid Chromatography-Tandem Mass Spectrometry Systems in Proteomics Analyses* , 2009, Molecular & Cellular Proteomics.

[41]  Henry H. N. Lam,et al.  Data analysis and bioinformatics tools for tandem mass spectrometry in proteomics. , 2008, Physiological genomics.

[42]  Benno Schwikowski,et al.  Alignment of LC‐MS images, with applications to biomarker discovery and protein identification , 2008, Proteomics.