Cleavable surfactants

• US 10,626,349 B2
• Filed: 10/10/2017
• Issued: 04/21/2020
• Est. Priority Date: 10/11/2007
• Status: Active Grant

• Current Assignee: Promega Corporation
• Original Assignee: Promega Corporation
• Inventors: Saveliev, Sergei, Simpson, Daniel, Wood, Keith V., Uyeda, Harry Tetsuo, Hitko, Carolyn Woodroofe, Klaubert, Dieter
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US15/729,406
• Publication Number:
  US 20180187127A1
• Time in Patent Office: 924 Days
• CPC Class Codes: C07C 309/10 with the oxygen atom of at least one of the etherified hydroxy groups further bound to an acyclic carbon atom
• CPC Class Codes: C07C 309/14 containing amino groups bound to the carbon skeleton
• CPC Class Codes: C07C 309/15 the nitrogen atom of at least one of the amino groups being part of any of the groups , X being a hetero atom, Y being any atom
• CPC Class Codes: C07C 309/16 containing doubly-bound nitrogen atoms bound to the carbon skeleton
• CPC Class Codes: C07C 323/12 the carbon skeleton being acyclic and saturated
• CPC Class Codes: C07C 329/06 having sulfur atoms of thiocarbonic groups bound to acyclic carbon atoms
• CPC Class Codes: C07D 307/40 Radicals substituted by oxygen atoms
• CPC Class Codes: C07D 307/56 with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
• CPC Class Codes: C07D 317/22 etherified
• CPC Class Codes: C07D 333/16 by oxygen atoms
• CPC Class Codes: C11D 1/143 Sulfonic acid esters
• CPC Class Codes: C11D 1/20 Fatty acid condensates

Abstract

The invention provides surfactant compounds of formulas I-IX, which can be used in methods for aiding the solubilization, digestion, preparation, analysis, and/or characterization of biological material, for example, proteins or cell membranes. The compounds can also aid in the recovery of peptides generated during protein digestion, particularly for in-gel digestion protocol. Additionally, the compounds can improve enzymatic protein deglycosylation without interfering with downstream sample preparation steps and mass spectrometric analysis. The compounds can be specifically useful as digestion aids that can be decomposed by an acid, by heat, or a combination thereof. Decomposition of the surfactants allows for facile separation from isolated samples, and/or allows for analysis of the sample without interfering with the sensitivity of various analytical techniques.

Claims

1. A method for protein digestion comprising contacting a gel, a solid support, or a solution, wherein the gel, the solid support, or the solution comprises a biomaterial that includes at least one protein, with a digestion reagent and a surfactant compound of formula (I), or a salt thereof, to provide a sample comprising at least one digested protein:

2. The method of claim 1 wherein the biomaterial is in a gel or is bound to a solid support.

3. The method of claim 1 wherein the biomaterial is in an aqueous solution.

4. The method claim 1 wherein the digestion reagent comprises one or more of a protease, CNBr, or hydroxylamine.

5. The method of claim 4 wherein the protease is a serine protease.

6. The method of claim 5 wherein the serine protease is trypsin or chymotrypsin.

7. The method of claim 1 further comprising isolating the one or more digested proteins.

8. The method of claim 1 further comprising analyzing the digested proteins, wherein the digested proteins are analyzed by mass spectrometry, liquid chromatography, gel electrophoresis, or a combination thereof.

9. The method of claim 1 wherein Q is (C₁-C₆)alkyl, phenyl, or phenyl-NH(C₁-C₆)alkyl.

10. The method of claim 1 wherein X is NH, and Z is O.

11. The method of claim 1 wherein M is H or Na.

12. The method of claim 1 wherein L is a direct bond.

13. The method of claim 1 wherein R¹ is (C₄-C₂₀)alkyl substituted with a (C₆-C₁₀)aryl or heteroaryl group.

14. The method of claim 1 wherein R¹ is (C₁₀-C₁₄)alkyl.

15. The method of claim 14 wherein R² and R³ are each (C₁-C₆)alkyl.

16. The method of claim 1 wherein the surfactant compound of formula (I) is

17. The method of claim 1 wherein the surfactant compound of formula (I) is

18. The method of claim 1 wherein the surfactant compound of formula (I) is

19. The method of claim 1, wherein the surfactant compound of formula (I) is a compound that is capable of degrading in the presence of an acid, heat, or a combination thereof.

20. The method of claim 19, wherein the method further comprising degrading the surfactant compound of formula (I) after protein digestion, wherein the degrading comprises contacting the surfactant compound with an acidic solution, heating the surfactant compound, or a combination thereof.

21. The method of claim 1, wherein the surfactant compound of formula (I) is a compound that is capable of self-hydrolyzing during protein digestion.