About iTop-Q

    Liquid chromatogram coupled with mass spectrometry (LC-MS) is a pre-dominant platform for proteomics research. Top-down proteomics using LC-MS has been increasingly applied for analyzing intact proteins to study genetic variation, alternative splicing, and post-translational modifications (PTMs) of the proteins, where each intact protein may have various distinct forms of variation or modification termed as proteoforms. However, only a few tools have been developed to perform charge state deconvolution, average molecular weight determination and quantitation of intact proteins from LC-MS spectra. Though Decon2LS and MASH Suite Pro have been available to provide intra-spectrum mass deconvolution and quantitation analysis, manual processing is still required to quantify proteoforms based on abundances obtained from multiple spectra. Currently there are few automated tools for large-scale proteoform quantitation based on inter-spectrum abundance in top-down proteomics. In this study, we present the first fully automated tool, called iTop-Q (intelligent Top-down proteomics Quantitation), for top-down proteomics quantitation using XIC abundances. Instead of utilizing single spectrum for proteoform quantitation, iTop-Q constructs extracted ion chromatograms (XICs) of possible proteoform peaks across adjacent MS1 spectra to calculate abundances for accurate quantitation. Notably, iTop-Q is implemented with a newly proposed algorithm, called DYAMOND, using dynamic programming for charge state deconvolution, instead of using THRASH as in other tools. Furthermore, iTop-Q performs proteoform alignment to support quantitation analysis across replicates/samples. The performance evaluation on an in-house standard data set and a public large-scale yeast lysate data set shows that iTop-Q achieves highly accurate quantitation, more consistent quantitation than using intra-spectrum quantitation. Furthermore, the DYAMOND algorithm is suitable for high charge state deconvolution and can distinguish shared peaks in co-eluting proteoforms.

 

The User Guide of iTop-Q(PDF)

Important Notices

  • For users using Windows Vista or Windows 7, please make sure that the "Privilege Level" of Compatibility has been checked. (Right click on the icon of iTop-Q->Compatibility->Privilege Level: Run this program as an administrator)

 

Tutorial Video

If you are unable to open the tutorial video, you may also download it (link).

 

System Requirement

  The recommended system environment is:
  1. Operating System: Windows XP, Windows 7, 8, 10 and Windows Server 2008, 2012
  2. CPU: Intel® CPU 2.40 GHz or faster
  3. Hard Disk: depending on the input file size. It would be safe to have the free disk space at least a few times the total size of input files.
  4. Memory: depending on the input file size. As a rule of thumb, 4GB or more is suggested. Larger input files will be more efficiently processed with larger free memory space.
  5. Microsoft .Net Framework: .Net 4.0 or higher
  6. Screen Resolution: 1024*768 or higher

Update Infromation

2017.02.16 V1.000 is available
  • iTop-Q is officially on-line.