Destructible surfactants and uses thereof
View Patent ↗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.
1. A method for enhancing chemical digestion of a biomolecule comprising contacting the biomolecule with (i) trypsin and (ii) 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 —OS0 3 − , —R 4 0S0 3 − , —R 4 0R 5 S0 3 − , and —OR 5 S0 3 − ,
wherein R 4 and R 5 are each, independently, lower alkyl; and
wherein the biomolecule is selected from the group consisting of a protein and a peptide, and
wherein the activity of said trypsin is maintained or increased upon contact with the surfactant;
thereby enhancing the chemical digestion of said biomolecule;
wherein the surfactant is degraded after the chemical digestion.
2. The method of claim 1 , wherein the chemical digestion is enhanced by accelerating the rate of chemical digestion of said biomolecule, increasing the yield of chemical digestion of said biomolecule or increasing the completeness of chemical digestion of said biomolecule or a combination thereof.
3. The method of claim 1 , wherein the activity of trypsin is maintained upon contact with the surfactant.
4. The method of claim 1 , wherein the activity of trypsin is increased upon contact with the surfactant.
5. The method of claim 3 or 4 , wherein the activity of trypsin is maintained or increased relative to the activity of trypsin in the presence of a surfactant other than the surfactant of-formula I.
6. The method of claim 5 , wherein the surfactant other than the surfactant of formula I is SDS.
7. The method of claim 1 , further comprising the step of analyzing the biomolecule following chemical digestion thereof.
8. The method of claim 1 , wherein the biomolecule is contained in a biological sample.
9. The method of claim 8 , 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.
10. The method of claim 1 , wherein the biomolecule is selected from the group consisting of a lipophilic protein, a receptor, a proteolytic protein, and a membrane-bound protein.
11. The method of claim 7 , wherein the step of analyzing the biomolecule comprises analysis 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.
12. The method of claim 11 , wherein the mass spectrometry is surface desorption ionization mass spectrometry.
13. The method of claim 7 , wherein the surfactant is degraded prior to analysis.
14. The method of claim 1 , wherein the trypsin is immobilized.
15. The method of claim 1 , wherein the biomolecule is selected from bovine serum albumin, lysozyme, ovalbumine, myoglobin, ubiquitin, and bacteriorhodopsin.
16. The method of claim 1 , wherein the surfactant is degraded by contact with an acidic solution.
17. 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.
18. The method of claim 1 wherein the surfactant has the following chemical structure:
19. The method of claim 1 wherein the surfactant has the following chemical structure:
20. The method of claim 1 wherein increasing the activity of trypsin facilitates on-line automation, separation, mass spectrometric analysis, or a combination thereof.
21. The method of claim 1 wherein increasing the activity of trypsin, is performed under microscale conditions.
22. The method of claim 1 wherein the digestion occurs in an electrophoretic gel.
23. The method of claim 1 wherein the digestion occurs in the presence one or more surfactants that are different from the surfactant in Formula I.
24. The method of claim 23 wherein the digestion occurs in the presence of SDS.
25. The method of claim 1 wherein the digestion occurs in the absence of SDS.
26. A kit for increasing the activity of trypsin for the chemical digestion of a biomolecule comprising:
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 —OS0 3 − , —R 4 0S0 3 − , —R 4 0R 5 S0 3 − , and —OR 5 S0 3 − ,
wherein R 4 and R 5 are each, independently, lower alkyl; and instructions for use.