METHODS, COMPOSITIONS, AND KITS FOR TRAPPING MODIFIED BIOMOLECULES
The present disclosure provides methods, compositions, and kits for trapping functionalized biomolecules. The methods, compositions, and kits utilize a trapping material comprising a reactive carbonyl group. The trapping material is designed to react rapidly and efficiently with a functionalized biomolecule that comprises a reactive amino group, thus removing the functionalized biomolecule from solution. The trapping material is therefore effective in the purification of target molecules after labeling reactions with an excess of the functionalized biomolecule.
1 . A method for trapping a biomolecule, comprising the step of:
contacting a solution comprising a functionalized biomolecule with a trapping material;
wherein the functionalized biomolecule comprises a reactive amino group; and
wherein the trapping material comprises a reactive carbonyl group.
2 . The method of claim 1 , wherein the solution further comprises a labeled target biomolecule.
3 . The method of claim 2 , wherein the target molecule is conjugated with the functionalized biomolecule.
4 . The method of claim 1 , wherein the functionalized biomolecule is a functionalized oligonucleotide, polypeptide, or polysaccharide.
5 . The method of claim 1 , wherein the reactive amino group is a hydrazino group or an oxyimino group.
6 . The method of claim 1 , wherein the reactive amino group reacts with the reactive carbonyl group to form a covalent linkage.
7 . The method of claim 6 , wherein the covalent linkage is a hydrazone or an oxime.
8 . The method of claim 1 , wherein the reactive carbonyl group is a reactive aldehyde group.
9 . The method of claim 1 , wherein the trapping material is a porous resin.
10 . The method of claim 1 , wherein the trapping material is a size-exclusion resin.
11 . The method of claim 1 , wherein the trapping material is an oxidized polysaccharide resin.
12 . The method of claim 1 , wherein the trapping material is prepared by the oxidation of a polysaccharide resin.
13 . The method of claim 1 , wherein the trapping material is a solid surface.
14 . The method of claim 1 , further comprising the step of separating the solution from the trapping material.
15 . The method of claim 14 , wherein the separating step comprises gravity flow separation, magnetic separation, or centrifugation.
16 . The method of claim 15 , wherein the separating step comprises centrifugation.
17 . The method of claim 1 , further comprising the step of reacting the functionalized biomolecule with a target molecule prior to contacting the solution comprising the functionalized biomolecule with the trapping material.
18 . The method of claim 17 , wherein the target molecule comprises a reactive carbonyl group.
19 . The method of claim 17 , further comprising the step of separating the solution from the trapping material.
20 . The method of claim 19 , wherein the separating step comprises gravity flow separation, magnetic separation, or centrifugation.
21 . The method of claim 20 , wherein the separating step comprises centrifugation.
22 .- 50 . (canceled)