Methods, Compositions, and Kits for Quantitating Antibodies

Abstract

The present teachings provide methods, compositions, and kits for quantitating antibodies. In some embodiments, the method comprises binding two proximity probes to two binding sites on the target antibody, wherein the proximity probes are comprised of a binding moiety and a thereto coupled oligonucleotide, wherein the binding moities of each proximity probe are the same, and wherein the binding sites for the binding moieties of the proximity probes are on one and the same target antibody. The binding moieties are allowed to bind to the target antibody and the oligonucleotides interact with each other if they are in close proximity to each other. Quantitating the degree of interaction between the oligonucleotides allows for quantitation of the target antibody.

1 . A method for quantitating a target antibody in solution, the method comprising; 
 binding two proximity probes to two binding sites on the target antibody, wherein the proximity probes are comprised of a binding moiety and a thereto coupled oligonucleotide, wherein the binding moities of each proximity probe are the same, and wherein the binding sites for the binding moieties of the proximity probes are on one and the same target antibody;    allowing the binding moieties to bind to the target antibody and allowing the oligonucleotide to interact with each other if they are in close proximity to each other; and,    quantitating the degree of interaction between the oligonucleotides.    
     
     
         2 . A method according to  claim 1  further comprising amplifying the interacted oligonucleotides to form an amplification product, and quantitating of amplification product.  
     
     
         3 . A method according to  claim 1 , wherein the binding moieties of the proximity probes are selected from a protein, such as a monoclonal or polyclonal antibody, lectin, soluble cell surface receptor, combinatorially derived protein from phage display or ribosome display, peptide, carbohydrate, nucleic acid, such as an aptamer, or combinations thereof.  
     
     
         4 . A method according to  claim 1 , wherein the interaction of the oligonucleotides coupled to the binding moieties is through hybridization to a common splint template and ligation of the oligonucleotides.  
     
     
         5 . A kit for quantitating one or more target antibodies in solution comprising; 
 a pair of proximity probes comprising binding moieties with affinity for the target antibody and each provided with an oligonucleotide capable of interacting with each other, wherein the binding moities of each proximity probe are the same, and wherein the binding sites for the binding moieties of the proximity probes are on one and the same target antibody.    
     
     
         6 . The kit according to  claim 5  further comprising a ligase and a splint template for joining the oligonucleotides.  
     
     
         7 . The kit according to  claim 5  further comprising PCR primers which correspond to each of the oligonucleotides.  
     
     
         8 . A reaction composition comprising; 
 two proximity probes bound to two binding sites on a target antibody, wherein the proximity probes are comprised of a binding moiety and a thereto coupled oligonucleotide, wherein the binding moities of each proximity probe are the same, and wherein the binding sites for the binding moieties of the proximity probes are on one and the same target antibody.    
     
     
         9 . The reaction composition according to  claim 8  further comprising a splint oligonucleotide, wherein the splint oligonucleotide is hybridized to the oligonucleotides of the proximity probes.