Biotin Derivatives

Abstract

Biotin derivatives, methods of using the biotin derivatives and kits comprising the biotin derivatives.

1 . A biotin derivative having structural formula (I): 
       
         
           
           
               
               
           
         
         wherein: 
         X is S or O; 
         R 1  is selected from H and a derivative group; 
         R 2  is selected from H and a derivative group; 
         Y is O or absent; 
         L is absent or is a linker; 
         R 3  is selected from —OR 4 , —COOR 4 , a reactive group, a protein, a peptide, an amino acid, a dextran, a monosaccharide, a disaccharide, a trisaccharide, an oligosaccharide, a polysaccharide, a hormone, a lipopolysaccharide, a nucleotide, an oligonucleotide, a small molecule, a cell, a microplate, and a microparticle; 
         R 4  is selected from H and a derivative group; 
         wherein at least one of R 1  and R 2  is H; 
         wherein if X is O, at least one of R 1  and R 2  is not H, or Y is O; 
         or structural formula (II): 
       
       
         
           
           
               
               
           
         
         wherein: 
         X is S or O; 
         R is selected from H and a derivative group; 
         R 2  is selected from H and a derivative group; 
         L is absent or is a linker; 
         R 3  is selected from —OR 4 , —COOR 4 , a reactive group, a protein, a peptide, an amino acid, a dextran, a monosaccharide, a disaccharide, a trisaccharide, an oligosaccharide, a polysaccharide, a hormone, a lipopolysaccharide, a nucleotide, an oligonucleotide, a small molecule, a cell, a microplate, and a microparticle; 
         R 4  is selected from H and a derivative group; 
         wherein at least one of R 1  and R 2  is H. 
       
     
     
         2 - 76 . (canceled) 
     
     
         77 . A composition comprising a biotin derivative of  claim 1  and a solvent. 
     
     
         78 . (canceled) 
     
     
         79 . A method of immobilizing a biotin derivative to a solid support, comprising contacting a solid support with a biotin derivative of  claim 1 , wherein the solid support comprises a biotin-binding moiety. 
     
     
         80 - 107 . (canceled) 
     
     
         108 . A method of detecting a protein in a sample, comprising the steps of:
 a. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group;   b. providing a sample comprising a protein comprising a complementary group capable of reacting with the reactive group;   c. providing a biotin-binding moiety comprising a detectable moiety;   d. contacting the biotin derivative with the sample to provide a labeling mixture;   e. contacting the labeling mixture with the biotin-binding moiety; and   f. detecting the protein.   
     
     
         109 - 114 . (canceled) 
     
     
         115 . A method of detecting a protein in a sample, comprising the steps of:
 a. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group; and wherein the linker is present and comprises a detectable moiety.   b. providing a sample comprising a protein comprising a complementary group capable of reacting with the reactive group;   c. contacting the biotin derivative with the sample; and   d. detecting the protein.   
     
     
         116 - 120 . (canceled) 
     
     
         121 . A method of isolating a protein in a sample, comprising the steps of:
 a. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group;   b. providing a sample comprising a protein comprising a complementary group capable of reacting with the reactive group;   c. providing a solid support comprising a biotin-binding moiety;   d. contacting the biotin derivative with the sample to provide a labeling mixture;   e. contacting the labeling mixture with the solid support; and   f. allowing the protein to bind to the solid support to provide the isolated protein.   
     
     
         122 - 126 . (canceled) 
     
     
         127 . A method of isolating a cell in a sample, comprising the steps of:
 a. providing a sample comprising at least one cell;   b. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group;   c. providing an antibody comprising a complementary group capable of reacting with the reactive group and capable of binding to a protein on the cell;   d. providing a solid support comprising a biotin-binding moiety;   e. contacting the biotin derivative with the antibody to provide a biotin-labeled antibody;   f. contacting the biotin-labeled antibody with the sample to provide a labeling mixture;   g. contacting the labeling mixture with the solid support; and   h. allowing the cell to bind to the solid support to provide the isolated cell.   
     
     
         128 - 132 . (canceled) 
     
     
         133 . A method of labeling a protein, comprising the steps of:
 a. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group;   b. providing a sample comprising a protein comprising a complementary group capable of reacting with the reactive group; and   c. contacting the biotin derivative with the sample to provide a biotin derivative-labeled protein.   
     
     
         134 - 137 . (canceled) 
     
     
         138 . The method of  claim 133 , further comprising the steps of:
 a. providing a biotin-binding moiety comprising a detectable moiety; and   b. contacting the biotin-binding moiety with the biotin derivative-labeled protein.   
     
     
         139 . The method of  claim 138 , wherein the biotin-binding moiety is selected from an avidin or a strepavidin. 
     
     
         140 . The method of  claim 138 , wherein the detectable moiety is a chromophore, a fluorescent dye, a fluorescent protein, a nanocrystal, an enzyme, a molecule comprising an atom which is a radioisotope, or an atom which is a radioisotope. 
     
     
         141 . A method of labeling a protein in a sample, comprising the steps of:
 a. providing a biotin derivative of  claim 1 , wherein R 3  is a reactive group; and wherein the linker is present and comprises a detectable moiety;   b. providing a sample comprising a protein comprising a complementary group capable of reacting with the reactive group; and   c. contacting the biotin derivative with the sample to provide a labeled protein.   
     
     
         142 . The method of  claim 141 , wherein the reactive group is an alkyne, and the complementary group is an azide. 
     
     
         143 . (canceled) 
     
     
         144 . The method of  claim 141 , wherein the reactive group is an azide and the complementary group is an alkyne. 
     
     
         145 . (canceled) 
     
     
         146 . The method of  claim 141 , wherein the detectable moiety is a chromophore, a fluorescent dye, a fluorescent protein, a nanocrystal, an enzyme, a molecule comprising an atom which is a radioisotope, or an atom which is a radioisotope. 
     
     
         147 . A method of labeling a biotin derivative comprising the steps of:
 a. providing a biotin derivative of  claim 1 ;   b. providing a biotin-binding moiety; and   c. contacting the biotin derivative with the biotin-binding moiety to provide the labeled biotin derivative.   
     
     
         148 - 149 . (canceled) 
     
     
         150 . A kit comprising a biotin derivative of  claim 1  and a biotin binding moiety. 
     
     
         151 . The kit of  claim 150 , wherein the biotin-binding moiety is selected from an avidin or a strepavidin. 
     
     
         152 . The kit of  claim 150 , wherein the biotin-binding moiety comprises a detectable moiety. 
     
     
         153 . The kit of claim  149 , wherein the detectable moiety is a chromophore, a fluorescent dye, a fluorescent protein, a nanocrystal, an enzyme, or a radioisotope.