Hello Shagun,

I downloaded the larger zip file twice and tried to decompress, but each
time it told me there was a corruption in the zip file.  The smaller files
downloaded fine.  Can you check your file and upload again?

Thanks!
-David

On Tue, Aug 13, 2024 at 9:47 AM Shagun Gupta <[email protected]> wrote:

> Hi David
>
> I have attached a link to the following files
> - one replicate per condition .d file (file starting with "C_" had IDs but
> no values that could be extracted for quantification)
> - comet parameter file I used
> - fasta file I ran the search with.
>
> Let me know if I can share anything else!
>
> Box_link_diagnosis
> <https://cornell.box.com/s/5y2uak2twszajaniaqpnevxn4zdneu54>
> Best
> Shagun
>
> On Tuesday, August 13, 2024 at 11:05:42 AM UTC-4 David Shteynberg wrote:
>
>> Hello Shagun,
>>
>> Thank you for the detailed report.  If you are able, please first
>> compress (into a zip or similar) and then share some of the problem .d
>> files so I can try to replicate this issue on my computer before I offer
>> any suggestions.
>>
>> Cheers!
>> -David
>>
>> On Tue, Aug 13, 2024 at 7:50 AM Shagun Gupta <[email protected]> wrote:
>>
>>> Hi all
>>>
>>> I have been experiencing issues processing .d files obtained from a
>>> Bruker timsTOF HT in DDA-LFQ mode, more specifically getting quantification
>>> - precursor intensity - per spectra. I am using TPP V6.3.3 Arcus on a
>>> windows computer.
>>>
>>> Details
>>> - There are 12 .d files (4 repeats of 3 conditions) composed of a human
>>> with yeast proteome spike in at different ratios.
>>> - Converted to .mzXML using msconvert
>>> - Searched with COMET and a search database taken from UniProt for Homo
>>> sapiens+Yeast
>>> - Processed with PeptideProphet (filtered at probability associated with
>>> 1% FDR), XPRESS, ProteinProphet. Ran with -PREC flag (PeptideProphet), -i
>>> flag (XPRESS)
>>> - Want to do hypothesis testing (comparing the three conditions
>>> pairwise) using MSstats. So require raw precursor intensity values to make
>>> a file that can be used as input to MSstats.
>>>
>>> Unfortunately after trying the above, and a few more things, while I get
>>> a large number of PSMs passing FDR (~30k), a large proportion of them do
>>> not have any precursor intensity value (<1k have some "light area" values).
>>> Using the "light area" values also does not give expected results (its a
>>> benchmarking dataset and processing with MSFragger gave excellent results
>>> that align with expected ratios etc.). Could you suggest things I could be
>>> doing differently to get the right results? (I imagine it might have
>>> something to do with the initial conversion to mzXML itself?)
>>>
>>> Happy to share any other details needed!
>>>
>>> Best
>>> Shagun
>>>
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