Dear David,
This is PeptideProphet issue (we don't use iProphet for this analysis). 
With the search parameters that I sent you followed by running the 
PeptideProphet command of *xinteract -Ninteract.pep.xml -p0.05 -l7 -PPM 
-OANEp -dDECOY Seq69478_QE2.pep.xml,*
and lastly using a cutoff of 1% error rate we obtain with TPP 5.2 2836 
correct assignments and with TPP 6.0 only 2228 correct assignments. The 
probability for 1% error rate is 0.84 in TPP 5.2 and 0.9090 in TPP 6.0).
In the following link, you will find the interact files (new and old):
https://www.dropbox.com/t/mwtpCLeEJRLCwbi1 
<https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.dropbox.com%2Fl%2FAACfc2b_KukHZAS1VuNo4sPM3gEXyXtDlPE&data=04%7C01%7Coklab%40technion.ac.il%7C342d2db781c2443416b108d8febfbc70%7Cf1502c4cee2e411c9715c855f6753b84%7C1%7C0%7C637539447557029368%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000&sdata=LBdAB3QsH9rUxvNKMwKjOKYDbQXFxma9dREKgS5siWQ%3D&reserved=0>
Thanks,
Oded



On Tuesday, 13 April 2021 at 03:38:33 UTC+3 David Shteynberg wrote:

> Dear Oded,  
>
> Thanks for this.  I ran a quick test and I actually observed a few more 
> PSMs for PeptideProphet 6.0.0-rc14 for the same PeptideProphet probability 
> cutoff for this dataset.  Is the issue you see with PeptideProphet or 
> iProphet results?   Which spectra were getting excluded in the your testing 
> of 5.2.0 vs 6.0.0, can you give me some specific examples? Are you certain 
> you are using identical analysis parameters for PeptideProphet and what are 
> those parameters? 
>
> Cheers, 
>
> David
>
> On Mon, Apr 12, 2021, 2:15 PM Oded <[email protected]> wrote:
>
>> And also the search results with the new vs the old version (although I 
>> think they are more or less the same): 
>> https://www.dropbox.com/t/8VyBVdeuoWdk982q
>>
>>
>> On Tuesday, 13 April 2021 at 00:09:05 UTC+3 Oded wrote:
>>
>>> Hi David,
>>> You can find the mzML, Comet parameters and FASTA file here:
>>> https://www.dropbox.com/t/Yld0ZBWhsuFajeLj
>>> The link is valid for 7 days.
>>> Many thanks,
>>> Oded
>>>
>>> On Monday, 12 April 2021 at 23:46:08 UTC+3 David Shteynberg wrote:
>>>
>>>> I don't mind taking a crack at it over vacation.   Please let me know 
>>>> where I can pull the data from. I might not have quick solution for you 
>>>> but 
>>>> I can get started looking for the problem.  Which search engine did you 
>>>> use 
>>>> here?   I would need mzML data search results and the fasta database to 
>>>> get 
>>>> started. 
>>>>
>>>> Thanks!
>>>>
>>>>
>>>>
>>>> Thanks!
>>>>
>>>> On Mon, Apr 12, 2021, 1:38 PM Oded <[email protected]> wrote:
>>>>
>>>>> Hi David,
>>>>> Thank you for the quick reply. We are using TPP v6.0.0-rc14 
>>>>> Noctilucent, Build 202103031119-8400 (Windows_NT-x86_64).
>>>>> As for the data let me know when you are back and I will transfer you 
>>>>> either the raw file or the search results.
>>>>> Enjoy your vacation.
>>>>> Thanks,
>>>>> Oded
>>>>> On Monday, 12 April 2021 at 22:17:53 UTC+3 David Shteynberg wrote:
>>>>>
>>>>>> Hi Oded,  Which release candidate are you referring to?  The earlier 
>>>>>> candidates may have a bug that is corrected in a later version. If you 
>>>>>> can 
>>>>>> share some data and specifics about the missing PSMs I can run it here 
>>>>>> and 
>>>>>> troubleshoot the problem.   
>>>>>>
>>>>>> Thanks!
>>>>>>
>>>>>> David 
>>>>>>
>>>>>> P.S. I am on vacation this week so will troubleshoot next week.
>>>>>>
>>>>>> On Mon, Apr 12, 2021, 11:49 AM Oded <[email protected]> wrote:
>>>>>>
>>>>>>> Hi there,
>>>>>>> We recently download the TPP 6.0 RC and while using it we noticed 
>>>>>>> that we obtain fewer peptides IDs than what we got for the same dataset 
>>>>>>> and 
>>>>>>> search output with version 5.2.
>>>>>>> Many of the missing peptides seem to have decent Expect value and 
>>>>>>> MS/MS following a visual inspection. 
>>>>>>> This seems that this is due to changes in PeptideProphet and are not 
>>>>>>> related to the search itself that was done with Comet.
>>>>>>> Is there any additional parameter that should be included in the new 
>>>>>>> version in order to restore the missing peptides?
>>>>>>> Many thanks,
>>>>>>> Oded
>>>>>>>
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