Destructible surfactants and uses thereof

Patent metadata

Field	Value
Publication number	US-9598716-B2
Application number	US-201314075637-A
Country	US
Kind code	B2
Filing date	Nov 8, 2013
Priority date	May 31, 2002
Publication date	Mar 21, 2017
Grant date	Mar 21, 2017

Abstract

The present invention provides methods for enhancing chemical reactions of molecules, e.g., biomolecules, with destructible surfactants. The chemical reactions may involve and/or be associate with analysis, e.g., solubilizing, separating, purifying and/or characterizing the molecules. In one aspect, the anionic surfactants of the present invention may be selectively broken up at relatively low pH. The resulting breakdown products of the surfactants may be removed from the molecule/sample with relative ease. The invention has applicability in a variety of analytical techniques.

First claim

What is claimed is:

1. A method for enhancing chemical digestion of a biomolecule comprising contacting the biomolecule with a protease, and a surfactant represented by formula I:

in which
p is 0, 1, or 2;
R is alkyl;
R 1 and R 2 are each, independently, hydrogen or methyl; and
R 3 is selected from —OSO 3 − , —R 4 OSO 3 − , —R 4 OR 5 SO 3 − ,
wherein R 4 and R 5 are each independently, lower alkyl;

to thereby enhance the chemical digestion of the biomolecule
wherein the ratio of the protease, to the total biomolecule is 1:100 or less.

2. The method of claim 1 , further comprising analyzing the biomolecule following the chemical reaction.

3. The method of claim 1 , wherein the biomolecule is contained in a biological sample.

4. The method of claim 3 , wherein the biological sample is selected from the group consisting of inclusion bodies, biological fluids, biological tissues, biological matrices, embedded tissue samples, and cell culture supernatants.

5. The method of claim 1 , wherein the biomolecule is selected from the group consisting of a protein and a peptide.

6. The method of claim 5 , wherein the biomolecule is selected from the group consisting of a lipophilic protein, a receptor, a proteolytic protein, and a membrane-bound protein.

7. The method of claim 2 , wherein the analysis is selected from the group consisting of solid phase extraction, solid phase micro extraction, electrophoresis, mass spectrometry, liquid chromatography, liquid-liquid extraction, membrane extraction, soxhlet extraction, precipitation, clarification, electrochemical detection, staining, elemental analysis, Edmund degradation, nuclear magnetic resonance, infrared analysis, flow injection analysis, capillary electrochromatography, ultraviolet detection, and combinations thereof.

8. The method of claim 7 , wherein the mass spectrometry is surface desorption ionization mass spectrometry.

9. The method of claim 8 , wherein the surfactant is not degraded prior to analysis.

10. The method of claim 8 , wherein the surfactant is degraded prior to analysis.

11. The method of claim 1 , further comprising separating the resulting biomolecule fragments.

12. The method of claim 1 , wherein the protease is immobilized.

13. The method of claim 1 , wherein the protease is selected from the group consisting Trypsin, Chymotrypsin, Lys-C, Staphylococcus aureus Protease V8, AspN, Arg-C, Clostripain, Pepsin, and Papain.

14. The method of claim 1 , further comprising degrading the surfactant after the chemical reaction.

15. The method of claim 14 , wherein the surfactant is degraded by contact with an acidic solution.

16. The method of claim 1 , wherein the surfactant is represented by formula II:

in which
R 6 is alkyl;
R 7 is selected from —OSO 3 − , —R 4 OSO 3 − , —R 4 OR 5 SO 3 − , and —OR 5 SO 3 − ,
wherein R 4 and R 5 are each, independently, lower alkyl.

17. The method of claim 1 wherein the surfactant has the following chemical structure:

18. The method of claim 1 wherein the surfactant has the following chemical structure:

19. The method of claim 1 wherein the digestion occurs in an electrophoretic gel.

20. The method of claim 19 wherein the digestion occurs in the presence one or more surfactants that are different from the surfactant in Formula I.

21. The method of claim 20 wherein the digestion occurs in the presence of SDS.

22. The method of claim 20 wherein the digestion occurs in the absence of SDS.

23. A method for analysis of a biomolecule comprising: enhancing chemical digestion of a biomolecule by contacting a sample containing the biomolecule with a protease, and a surfactant represented by formula I:

in which
p is 0, 1, or 2;
R is alkyl;
R 1 and R 2 are each, independently, hydrogen or methyl; and
R 3 is selected from —OSO 3 − , —R 4 OSO 3 − , —R 4 OR 5 SO 3 − ,
wherein R 4 and R 5 are each independently, lower alkyl;

to thereby enhance the chemical digestion of the biomolecule
wherein the ratio of the protease, to the total biomolecule is 1:100 or less.

24. The method of claim 23 , wherein the molecule is a biomolecule.

25. The method of claim 24 , further comprising analyzing the biomolecule following the chemical reaction.

26. The method of claim 24 , wherein the biomolecule is contained in a biological sample.

27. The method of claim 26 , wherein the biological sample is selected from the group consisting of inclusion bodies, biological fluids, biological tissues, biological matrices, embedded tissue samples, and cell culture supernatants.

28. The method of claim 24 , wherein the biomolecule is selected from the group consisting of a protein and a peptide.

29. The method of claim 28 , wherein the biomolecule is selected from the group consisting of a lipophilic protein, a receptor, a proteolytic protein, and a membrane-bound protein.

30. The method of claim 25 , wherein the analysis is selected from the group consisting of solid phase extraction, solid phase micro extraction, electrophoresis, mass spectrometry, liquid chromatography, liquid-liquid extraction, membrane extraction, soxhlet extraction, precipitation, clarification, electrochemical detection, staining, elemental analysis, Edmund degradation, nuclear magnetic resonance, infrared analysis, flow injection analysis, capillary electrochromatography, ultraviolet detection, and combinations thereof.

31. The method of claim 30 , wherein the mass spectrometry is surface desorption ionization mass spectrometry.

32. The method of claim 31 , wherein the surfactant is not degraded prior to analysis.

33. The method of claim 23 wherein the digestion occurs in an electrophoretic gel.

34. The method of claim 33 wherein the digestion occurs in the presence of SDS.

35. The method of claim 33 wherein the digestion occurs in the absence of SDS.

Assignees

• Waters Technologies Corp

Inventors

• Bouvier Edouard S
• Compton Bruce J
• Gebler John C
• Gilar Martin
• Yu Ying Q
• Lee Peter J
• Kane Elizabeth K

Classifications

• G01N33/5306

  Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding · CPC title

• G01N27/44726

  using specific dyes, markers or binding molecules · CPC title

• C12P21/06 Primary

  produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00) · CPC title

• G01N27/44747

  Composition of gel or of carrier mixture · CPC title

• C12Q1/37 Primary

  involving peptidase or proteinase · CPC title