Quality control reagents and methods

Patent metadata

Field	Value
Publication number	US-2021156832-A1
Application number	US-202117159923-A
Country	US
Kind code	A1
Filing date	Jan 27, 2021
Priority date	Feb 13, 2013
Publication date	May 27, 2021
Grant date	—

Abstract

The present invention provides reagents for instrumentation quality control and methods of use thereof. In particular, sets of peptides or other molecules are provided for evaluating the performance of instruments with mass spectrometry (MS) and/or liquid chromatography (LC) functionalities.

First claim

1 . A peptide mixture comprising two or more distinct-mass versions of each of two or more distinct-sequence peptides.

2 . The peptide mixture of claim 1 , wherein each of the distinct-sequence peptides is of distinct hydrophobicity.

3 . The peptide mixture of claim 1 , wherein all of the distinct-mass versions of any of the distinct-sequence peptides are present at distinct concentrations.

4 . The peptide mixture of claim 3 , wherein the distinct-mass versions of any of the distinct-sequence peptides are present at concentrations ranging from at least as low as 10 nM to at least as high at 1 μM.

5 . The peptide mixture of claim 4 , wherein the distinct-mass versions of any of the distinct-sequence peptides are present at concentrations ranging from 1 nM (femtomoles per microliter) to 10 μM.

6 . The peptide mixture of claim 1 , wherein the distinct-sequence peptides are separable by liquid chromatography based on their different hydrophobicities.

7 . The peptide mixture of claim 1 , wherein the distinct-sequence peptides are separable by liquid chromatography based on their different charge, size, or hydrophilicity.

8 . The peptide mixture of claim 1 , comprising 3-20 distinct-sequence peptides.

9 . The peptide mixture of claim 8 , comprising 5-10 distinct-sequence peptides.

10 . The peptide mixture of claim 1 , wherein the distinct-mass versions of any of the distinct-sequence peptides are differentiable by mass spectrometry.

11 . The peptide mixture of claim 1 , wherein the distinct-mass versions of any of the distinct-sequence peptides are the result of different combinations of stable heavy isotope-labeled amino acids.

12 . The peptide mixture of claim 11 , wherein each of the distinct-mass versions of any of the distinct-sequence peptides comprises a different number of uniformly stable isotope-labeled amino acids.

13 . The peptide mixture of claim 1 , comprising 3-20 distinct-mass versions of each of the distinct-sequence peptides.

14 . The peptide mixture of claim 13 , comprising 5-10 distinct-mass versions of each of the distinct-sequence peptides.

15 . A method for assessing performance of an instrument with both liquid chromatography (LC) and mass spectrometry (MS) functionalities comprising:
(a) introducing a peptide mixture to the instrument, wherein said peptide mixture comprises two or more distinct-mass versions of each of two or more distinct-sequence peptides; (b) separating or analyzing the peptide mixture by LC; (c) analyzing the peptide mixture by MS; and (d) assessing the performance of the LC and MS functionalities of the instrument based on results of steps (b) and (c).

16 . The method of claim 15 , wherein the peptide mixture purified peptides.

17 . The method of claim 15 , wherein the peptide mixture comprises peptides in the presence of one or more impurities.

18 . The method of claim 15 , wherein each distinct-mass version of a distinct-sequence peptide is present at a different concentration.

19 . The method of claim 15 , wherein assessing the performance of the LC and MS functionalities of the instrument comprises reporting one or more LC-parameters for each peptide sequence selected from: retention times, peak height, peak width, peak resolution, and peak symmetry.

20 . The method of claim 15 , wherein assessing the performance of the LC and MS functionalities of the instrument comprises reporting one or more MS-parameters for each distinctly-massed versions of one or more of the peptide sequences selected from: resolution, mass accuracy, sensitivity of “neat” samples, sensitivity of complex samples, dynamic range, mass resolution, isolation efficiency, MS/MS spectral quality and linear response in a single analytical experiment.

21 . The method of claim 15 , wherein the sensitivity of LC and MS functionalities

22 . The method of claim 15 , wherein the assessment of the performance of the LC and MS functionalities is performed by software.

23 . The method of claim 22 , wherein the software generates a performance score.

24 . The method of claim 15 , wherein said distinct-sequence peptides are of distinct hydrophobicities and separable by LC.

25 . The method of claim 24 , wherein said distinct-mass versions comprise different combinations of heavy isotope labeled amino acids and is separable by MS.

Assignees

• Promega Corp

Inventors

• Coon Joshua Jacques
• Rosenblatt Michael M
• Urh Marjeta

Classifications

• G01N2030/8831

  involving peptides or proteins · CPC title

• G01N30/8665 Primary

  for calibrating the measuring apparatus · CPC title

• G01N2030/047

  external · CPC title

• H01J49/0009

  Calibration of the apparatus · CPC title

• G01N2030/027

  Liquid chromatography · CPC title