Multi-amplitude modular labeled compounds

Abstract

Sets of compounds bearing detectably different groups of labels are provided. Typically, different compounds bear different numbers of a single type of label and are thus distinguishable by the amplitude of signal produced by the label. The compounds are assembled from label components and protein cores to facilitate modular production of the compounds. In compounds containing two or more proteins, the proteins are typically covalently linked. Useful sets of compounds include sets of labeled nucleotide analogs, particularly dye-label nucleotide analogs that include tetravalent biotin-binding protein cores.

1 - 49 . (canceled) 
     
     
         50 . A composition comprising:
 a first compound comprising one or more tetravalent biotin-binding proteins and one or more label components bound to the one or more tetravalent biotin-binding proteins, each label component comprising one or more labels; and   a second compound comprising two or more covalently linked tetravalent biotin-binding proteins and two or more label components bound to the two or more tetravalent biotin-binding proteins, each label component comprising one or more labels;   wherein the total number of labels in the second compound is greater than that in the first compound.   
     
     
         51 . The composition of  claim 50 , wherein the first compound comprises two or more covalently linked tetravalent biotin-binding proteins. 
     
     
         52 . The composition of  claim 50 , wherein the label components on the first and second compounds are identical. 
     
     
         53 . The composition of  claim 50 , wherein the labels are all identical. 
     
     
         54 - 55 . (canceled) 
     
     
         56 . The composition of  claim 50 , wherein each label component is bound to the tetrameric biotin-binding protein through a biotin moiety. 
     
     
         57 . The composition of  claim 50 , wherein each label component comprises a bis-biotin moiety bound to two biotin binding sites on one of the tetravalent biotin-binding proteins. 
     
     
         58 . The composition of  claim 50 , wherein the one or more tetravalent biotin-binding proteins in the first compound and the two or more tetravalent biotin-binding proteins in the second compound comprise streptavidin or traptavidin. 
     
     
         59 . (canceled) 
     
     
         60 . The composition of  claim 50 , comprising a third compound comprising two or more covalently linked tetravalent biotin-binding proteins and three or more label components bound to the three or more tetravalent biotin-binding proteins, each label component comprising one or more labels;
 wherein the total number of labels in the third compound is greater than that in the second compound.   
     
     
         61 . The composition of  claim 60 , comprising a fourth compound comprising three or more covalently linked tetravalent biotin-binding proteins and four or more label components bound to the three or more tetravalent biotin-binding proteins, each label component comprising one or more labels;
 wherein the total number of labels in the fourth compound is greater than that in the third compound.   
     
     
         62 . The composition of  claim 61 , wherein the first compound comprises one label component, the second compound comprises two label components, the third compound comprises three label components, and the fourth compound comprises four label components, wherein each label component comprises a single, identical label. 
     
     
         63 . (canceled) 
     
     
         64 . The composition of  claim 61 , wherein the first compound comprises one tetravalent biotin-binding protein and one label component; wherein the second compound comprises two covalently linked tetravalent biotin-binding proteins and two label components; wherein the third compound comprises two covalently linked tetravalent biotin-binding proteins and three label components; and wherein the fourth compound comprises three covalently linked tetravalent biotin-binding proteins and four label components. 
     
     
         65 . (canceled) 
     
     
         66 . The composition of  claim 50 , wherein the labels are optical labels. 
     
     
         67 . The composition of  claim 66 , wherein the labels are fluorescent dye moieties. 
     
     
         68 . The composition of  claim 50 , wherein the second compound comprises two or more tetravalent biotin-binding proteins covalently linked by at least one isopeptide bond. 
     
     
         69 . (canceled) 
     
     
         70 . The composition of  claim 50 , wherein the second compound comprises two or more tetravalent biotin-binding proteins covalently linked through at least one bifunctional or multifunctional crosslinker. 
     
     
         71 . (canceled) 
     
     
         72 . The composition of  claim 50 , wherein the first compound is bound to at least one first molecule of interest, and the second compound is bound to at least one second molecule of interest. 
     
     
         73 . The composition of  claim 72 , wherein the first molecule of interest is a first antibody specific for a first epitope, the second molecule of interest is a second antibody specific for a second epitope, and the first and second epitopes are different. 
     
     
         74 . The composition of  claim 72 , wherein the first molecule of interest is a first nucleotide and the second molecule of interest is a second nucleotide different from the first. 
     
     
         75 . (canceled) 
     
     
         76 . The composition of  claim 72 , wherein each molecule of interest comprises a bis-biotin moiety bound to two biotin binding sites on one of the tetravalent biotin-binding proteins. 
     
     
         77 - 78 . (canceled) 
     
     
         79 . The composition of  claim 72 , wherein the first compound comprises two or more covalently linked tetravalent biotin-binding proteins and two or more first molecules of interest bound to the proteins.