Nuclease inhibitors and methods for their use
A class of anionic oligomers and polymers that function for inhibition of nucleases, particularly RNase. Specific inhibitors include mixtures of oligomers of vinyl sulfate. Methods for inhibition or inactivation of one or more nucleases in vitro which comprises the step of contacting the one or more nucleases in a biological medium with one or more of the anionic oligomeric or polymeric inhibitors of this invention. Kits for carrying out a biological procedure, biological reaction and/or a biological assay containing one or more inhibitors of this invention. The use of oligomers and/or polymers of this invention as additives in buffers or reagents. The inhibitors of the invention can also be attached to surfaces to provide for removal of nucleases from media, solutions or other liquids in contact with the solid.
1. A method for inhibiting one or more nuclease in vitro which comprises the step of contacting the one or more nuclease with an inhibitory composition which comprises one or more of the oligomers, polymers or both of formula:
where X is —SO 3 , —OSO 3 —PO 3 H, or —OPO 3 H, or salts thereof, n is an integer representing the number of repeating monomer units and n is 8 to 19, wherein the one or more oligomers, polymers or both are covalently bound to a solid.
2. The method of claim 1 in a biological medium wherein the contacting step comprises the step of adding the inhibitory composition to the biological medium.
3. The method of claim 1 wherein X is —SO 3 − .
4. The method of claim 3 wherein n is 8 to 18.
5. The method of claim 3 wherein n is 9 to 17.
6. The method of claim 1 wherein X is —SO 3 − .
7. The method of claim 6 wherein n is 8 to 18.
8. The method of claim 6 wherein n is 9 to 17.
9. The method of claim 1 wherein the nuclease is an RNase.
10. The method of claim 1 which comprises the step of contacting the biological medium with a material of formula:
where X is —SO 3 , —OSO 3 —PO 3 H, or —OPO 3 H, or salts thereof, where n is 8 to 19, L is a linker or spacer group and S is a solid.
11. The method of claim 10 wherein X is —SO 3 − .
12. The method of claim 10 wherein the solid is silica gel.
13. The method of claim 10 wherein the solid is a glass surface.
14. The method of claim 10 wherein the linker is —(CH 2 )p-NH—CO—C(CH 3 )— where p is 1 or more.
15. The method of claim 14 wherein p is 3-10.
16. The method of claim 14 where p is greater than 10.
17. The method of claim 1 wherein the solid is silica gel particles, glass beads, glass surfaces, cross-linked polystyrene, cross-linked PEG, microspheres, plastic surfaces or plastic beads.
18. The method of claim 1 wherein the solid is the internal surface of a container in which a buffer or reagent is to be carried or stored or in which a procedure, purification, isolation, assay or reaction is to be carried out.
19. The method of claim 1 wherein the inhibitory composition is a mixture of one or more oligomers covalently bound to the solid.
20. The method of claim 10 wherein the solid is silica gel particles, glass beads, glass surfaces, cross-linked polystyrene, cross-linked PEG, microspheres, plastic surfaces or plastic beads.
21. The method of claim 10 wherein n is 8 to 18.
22. The method of claim 1 wherein n is 8 to 18.
23. The method of claim 10 wherein n is 9 to 17.
24. The method of claim 1 wherein the oligomer or polymer is covalently bonded to the solid by polymerization of monomers of the oligomer or polymer on the surface.
25. The method of claim 10 wherein the oligomer or polymer is covalently bonded to the solid by polymerization of monomers of the oligomer or polymer on linker L.
26. The method of claim 11 wherein n is 8 to 18.
27. The method of claim 10 wherein the nuclease is a RNase.
28. A method for inhibiting one or more nuclease in vitro which comprises the step of contacting the one or more nuclease with an inhibitory composition which comprises one or more of the oligomers, polymers or both of formula:
where X is —SO 3 , —OSO 3 —PO 3 H, or —OPO 3 H, or salts thereof, n is an integer representing the number of repeating monomer units and n is 8 to 19.
29. The method of claim 28 wherein n is 8 to 18.
30. The method of claim 28 wherein X is —SO 3 − .
31. The method of claim 30 wherein n is 8 to 18.
32. The method of claim 30 wherein n is 9-17.
33. The method of claim 1 where n is 9 to 17.