Quality control reagents and methods

Patent metadata

Field	Value
Publication number	US-10900939-B2
Application number	US-201916275417-A
Country	US
Kind code	B2
Filing date	Feb 14, 2019
Priority date	Feb 13, 2013
Publication date	Jan 26, 2021
Grant date	Jan 26, 2021

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

The invention claimed is:

1. A method for assessing performance of an instrument with both liquid chromatography (LC) and mass spectrometry (MS) functionalities comprising:
(a) introducing a neat peptide mixture in the absence of other background causing peptides into the instrument, wherein the neat peptide mixture comprises three or more distinct-mass isotopologues of each of three or more distinct peptide sequences, wherein two or more of the distinct-mass isotopologues of each of the three or more distinct peptide sequences comprises one or more amino acids with above natural abundance levels of one or more heavy isotopes;
(b) separating or analyzing the peptide mixture by LC; and
(c) analyzing the peptide mixture by MS;
wherein there is a linear relationship between the relative MS intensity versus the molar amount of the isotopologues for each distinct peptide sequence.

2. The method of claim 1 , wherein each of the distinct peptide sequences is of distinct hydrophobicity.

3. The method of claim 1 , wherein all of the distinct-mass isotopologues of any of the distinct peptide sequences are present at distinct concentrations.

4. The method of claim 1 , wherein the distinct-mass isotopologues of any of the distinct peptide sequences are present at concentrations ranging from at least as low as 1 nM to at least as high at 10 μM.

5. The method of claim 4 , wherein the distinct-mass isotopologues of any of the distinct peptide sequences are present at concentrations ranging from at least as low as 10 nM to at least as high at 1 μM.

6. The method of claim 1 , wherein the distinct peptide sequences are separable by liquid chromatography based on their different hydrophobicities.

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

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

9. The method of claim 8 , comprising 5-10 distinct peptide sequences.

10. The method of claim 1 , wherein the distinct-mass isotopologues of any of the distinct peptide sequences are differentiable by mass spectrometry.

11. The method of claim 1 , wherein the distinct-mass isotopologues of any of the distinct peptide sequences are the result of different combinations of stable heavy isotope-labeled amino acids.

12. The method of claim 1 , wherein each of the distinct-mass isotopologues of any of the distinct peptide sequences comprises a different number of uniformly stable isotope-labeled amino acids.

13. The method of claim 1 , comprising 3-20 distinct-mass isotopologues of each of the distinct peptide sequences.

14. The method of claim 13 , comprising 5-10 distinct-mass isotopologues of each of the distinct peptide sequences.

15. The method of claim 1 , wherein MS is MS/MS.

16. The method of claim 1 , further comprising:
(d) assessing the performance of the LC and MS functionalities of the instrument based on the analyses of steps (b) and (c).

17. The method of claim 1 , further comprising:
(d) repeating steps (a)-(c) at a second time point;
(e) comparing results of steps (b)-(c) at the first and second time points; and
(f) assessing the performance of the LC and MS functionalities of the instrument over time based on the comparison of step (e).

18. A method for determining the dynamic range of an instrument with both liquid chromatography (LC) and mass spectrometry (MS) functionalities comprising:
(a) introducing a neat peptide mixture in the absence of other background causing peptides into the instrument, wherein the neat peptide mixture comprises three or more distinct-mass isotopologues of each of three or more distinct peptide sequences wherein two or more of the distinct-mass isotopologues of each of the three or more distinct peptide sequences comprises one or more amino acids with above natural abundance levels of one or more heavy isotopes;
(b) separating or analyzing the peptide mixture by LC; and
(c) analyzing the peptide mixture by MS;
wherein the dynamic range of the instrument extends over the range for which there is a linear relationship between the relative MS intensity versus the molar amount of the isotopologues for each distinct peptide sequence.

Assignees

• Promega Corp

Inventors

• Coon Joshua Jacques
• Rosenblatt Michael M
• Urh Marjeta

Classifications

• G01N2030/8831

  involving peptides or proteins · CPC title

• H01J49/0009

  Calibration of the apparatus · CPC title

• G01N30/8665 Primary

  for calibrating the measuring apparatus · CPC title

• G01N2030/047

  external · CPC title

• G01N2030/027

  Liquid chromatography · CPC title