Compositions for analyzing proteins
View Patent ↗Methods, compositions and kits are disclosed for determining one or more target polypeptides in a sample where the target polypeptides have undergone a post-translational modification.
1. A compound of the formula:
T-L-M-D,
wherein:
T is a polypeptide-binding moiety;
L is
wherein * indicates the point of attachment to M, and ** indicates the point of attachment to T;
M is a divalent moiety comprising up to 100 non-hydrogen atoms selected from the group consisting of carbon, nitrogen, oxygen, phosphorus, boron, and sulfur, wherein M contains at least a group having the structure
D is a fluorescent dye selected from the group consisting of a water-soluble rhodamine dye, a fluorescein, and a 4,7-dichlorofluorescein;
and wherein *** indicates the point of attachment directly to an sp2-hybridized carbon atom of the non-fused benzene ring of the fluorescent dye.
2. The compound of claim 1 , wherein T comprises an antibody or a fragment thereof.
3. The compound of claim 1 , wherein T comprises an antibody specific for a target protein or polypeptide.
4. The compound of claim 1 , wherein T comprises a haptenized antibody.
5. The compound of claim 1 , wherein T comprises a biotinylated protein.
6. The compound of claim 1 , wherein T comprises an antibody derivatized with a functional polymer.
7. The compound of claim 1 , wherein M comprises a positively charged group.
8. The compound of claim 1 , wherein M comprises a negatively charged group.
9. The compound of claim 1 , wherein M comprises more than one amide group.
10. The compound of claim 9 , wherein M is
where *** indicates the point of attachment to D, and * indicates the point of attachment to L.
11. The compound of claim 1 , wherein D is a flourescein, where the fluorescein is selected from the group consisting of: 5-carboxyfluorescein, 6-carboxyfluorescein, 5-carboxy-4,7-dichlorofluorescein, 6-carboxy-4,7-dichlorofluorescein, 2′,7′-dimethoxy-5-carboxy-4,7-dichlorofluorescein, 2′,7′-dimethoxy-6-carboxy-4,7-dichlorofluorescein, 2′,7′dimethoxy-4′,5′-dichloro-5-carboxyfluorescein, 2′,7′dimethoxy-4′,5′-dichloro-6-carboxyfluorescein, 2′,7′-dimethoxy-4′,5′-dichloro-5-carboxy-4,7-dichlorofluorescein, 2′,7′ dimethoxy-4′,5′-dichloro-6-carboxy-4,7-dichlorofluorescein, 1′,2′,7′,8′-dibenzo-5-carboxy-4,7-dichlorofluorescein, 1′,2′,7′,8′-dibenzo-6-carboxy-4,7-dichlorofluorescein, 1′,2′,7′,8′-dibenzo-4′,5′-dichloro-5-carboxy-4,7-dichlorofluorescein, 1′,2′,7′,8′-dibenzo-4′,5′-dichloro-6-carboxy-4,7-dichlorofluorescein, 2′,7′-dichloro-5-carboxy-4,7-dichlorofluorescein, 2′,7′-dichloro-6-carboxy-4,7-dichlorofluorescein, 2′,4′,5′,7′-tetrachloro-5-carboxy-4,7dichlorofluorescein, and 2′,4′,5′,7′-tetrachloro-6-carboxy-4,7dichlorofluorescein.
12. The compound of claim 1 , wherein D is
and *** indicates the point of attachment to M.