Alkyl-linked nucleotide compositions
AssigneeSerenex, Inc.
InventorsKlass P. Hardeman, Steven E. Hall, Roy W. Ware, Lindsay A. Hinkley, Matthew G. Jenks
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Abstract
Alkyl-linked nucleotide non-homogeneous solid supports and nucleotide affinity media comprising an alkyl-linked nucleotide are provided. The linker is generally a hydrophobic linker that can be a 3, 4, 5, 6, 7, 8, 9, 10, or a longer carbon chain. Also included in the invention are methods for synthesis of an alkyl-linked nucleotide, nucleotide affinity media and methods of use thereof for affinity chromatography and screening methods.
Claims12
An alkyl-linked nucleotide non-homogeneous solid support consisting essentially of the general formula:
- wherein Y is a solid support;
- x=1;
- R1 is a covalent bond between Y and R2, or R1 is a divalent acyl group —C(═Q)—, wherein Q is O or NH2+;
- K is NH;
- R7 is (P)n where P is a phosphate or a thiophosphate and n is at least one;
- and m is at least one;
- Z is a 5′-nucleosidyl group or a 5′-nucleosidyl group wherein the 5′-nucleosidyl group is not naturally occurring, or a derivative thereof;
- and —R2—K— is selected from the group consisting of:
- wherein the solid support has a loading of an alkyl-linked nucleotide having a range of about 20% to about 50%.
A method for synthesizing a nucleotide affinity medium consisting essentially of the general formula:
- comprising the steps of:
- a) coupling at least one linker to a solid support in a suitable coupling buffer, wherein said linker is R2 or a combination of R1 and R2;
- b) end-capping at least a portion of reactive sites remaining on said solid support after said coupling step;
- and
- c) reacting a terminal phosphate or thiophosphate group of a nucleotide with said linker coupled to said solid support,
- wherein Y is a solid support;
- x=1;
- R1 is a covalent bond between Y and R2, or R1 is a divalent acyl group —C(═Q)—, wherein Q is O or NH2+;
- K is NH;
- R7 is (P)n where P is a phosphate or thiophosphate and n is at least one;
- Z is a 5′-nucleosidyl group or a 5′-nucleosidyl group wherein the 5′-nucleosidyl group is not naturally occurring, or a derivative thereof;
- and m is at least one;
- and —R2—K— is selected from the group consisting of:
- wherein the solid support has a loading of an alkyl-linked nucleotide having a range of about 20% to about 50%.
A method for screening a test compound comprising the steps of:
- a) contacting a proteome with a nucleotide affinity medium consisting essentially of the general formula:
- wherein Y is a solid support;
- x=1;
- R1 is a covalent bond between Y and R2, or R1 is a divalent acyl group —C(═Q)—, wherein Q is O or NH2+, K is NH;
- R7 is (P)n where P is a phosphate or thiophosphate and n is at least one Z is a 5′-nucleosidyl group or a 5′-nucleosidyl group wherein the nucleoside is not naturally occurring, or a derivative thereof;
- and m is at least one;
- and —R2—K— is selected from the group consisting of:
- wherein the solid support has a loading of an alkyl-linked nucleotide having a range of about 20% to about 50%;
- b) washing the nucleotide affinity medium with a buffer, whereby non-specifically bound components of the proteome are eluted from the nucleotide affinity medium and specific components of the proteome remain bound to the nucleotide affinity medium;
- c) contacting the nucleotide affinity medium bound with specific components of the proteome with at least one test compound;
- d) eluting from the nucleotide affinity medium components of the proteome that are specifically displaced by the test compound;
- and
- e) identifying the components of the proteome that are specifically displaced by the test compound from the nucleotide affinity medium.
Dependencies are inferred from the claim text.