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 soled support which comprises a mixture of oligomers of formula:
where X is —SO 3 − , —OSO 3 − , —PO 3 H − , or —OPO 3 H − , or salts thereof immobilized on the solid support and n is an integer representing the number of repeating monomer units and n is 8 to 19.
2. The solid support of claim 1 wherein n is 8 to 18.
3. The soled support of claim 1 wherein x is —SO 3 − .
4. The solid support of claim 3 wherein n is 8 to 18.
5. The solid support of claim 1 where the mixture of oligomers is immobilized on the solid support by sequential polymerization of monomers.
6. The solid support of claim 1 wherein the mixture of oligomers is immobilized on the solid support by radical polymerization of vinyl sulfonate.
7. The solid support of claim 1 which is selected from the group consisting of silica gel particles, glass beads, glass surfaces, cross-linked polystyrene, cross-linked polyethylene glycol, microspheres, plastic surfaces and plastic beads.
8. The solid support of claim 1 wherein the immobilized mixture of oligomers is represented by formula:
where X is —SO 3 − , —OSO 3 − , —PO 3 H − , or —OPO 3 H − , or salts, L is a linker bonded to the solid support and n is an integer representing the number of repeating monomer units and n is 8 to 19.
9. The solid support of claim 8 wherein n is 8 to 18.
10. The solid support of claim 8 wherein X is —SO 3 − .
11. The solid support of claim 10 wherein n is 8 to 18.
12. The solid support of claim 8 wherein the linker group is —(CH 2 ) 3 —NH—CO—CH(CH 3 )—.
13. The solid support of claim 8 which is selected from the group consisting of silica gel particles, glass beads, glass surfaces, cross-linked polystyrene, cross-linked polyethylene glycol, microspheres, plastic surfaces and plastic beads.
14. The solid support of claim 8 wherein the solid is derivatized with the linker group and the mixture of oligomers is immobilized on the solid support by sequential polymerization of monomers onto the linker group.
15. The solid support of claim 14 wherein the mixture of oligomers is immobilized on the solid support by radical polymerization of vinyl sulfonate.
16. A kit for a carrying out a procedure, assay or reaction in which the presence of a nuclease would be detrimental which comprises one or more solid supports of claim 1 .
17. The solid support of claim 3 wherein n is 9 to 17.
18. The solid support of claim 1 wherein n is 9 to 17.
19. The solid support of claim 8 wherein n is 9 to 17.
20. The solid support of claim 10 wherein n is 9 to 17.