Immunoassay and reagents and kits for performing the same

Abstract

A sandwich immunoassay is disclosed that provides simple to perform yet sensitive identification of analytes in samples. All assay constituents needed (except analyte to be detected) for one assay are dried. Upon reconstitution with sample, a 10 to 15 minute incubation gives a rapid and convenient detection assay capability. The method incorporates the capture of antigen to an immobilized capture antibody. A labeled reporter antibody with the molecule, binds to the antigen to form an immunocomplex capable of generating a detectable signal.

1 - 14 . (canceled)  
   
   
       15 : A kit, comprising: 
 (A) a container; and    (B) a reagent mixture comprising: 
 (1) an immobilized capture antibody; and  
 (2) a labeled reporter antibody,  
 wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a liquid comprising said labeled reporter antibody in the presence of said immobilized capture antibody, and wherein said reagent mixture is contained in said container, and  
   wherein said container is a tube which is capable of being hermetically sealed and wherein said tube comprises an indentation near the open end.    
   
   
       16 : The kit of  claim 15 , wherein said capture antibody is immobilized on a paramagnetic bead.  
   
   
       17 : The kit of  claim 15 , wherein said drying is carried out by lyophilization.  
   
   
       18 : The kit of  claim 15 , wherein said labeled reporter antibody is labeled with a tris bipyridyl ruthenium group.  
   
   
       19 . The kit of  claim 15 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       20 : The kit of  claim 15 , wherein said tube is a polypropylene tube.  
   
   
       21 : The kit of  claim 15 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       22 : The kit of  claim 15 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       23 : The kit of  claim 15 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.  
   
   
       24 : A method for detecting or quantifying an analyte, comprising: 
 (i) providing a kit comprising: 
 (A) a container; and  
 (B) a reagent mixture comprising: 
 (1) an immobilized capture antibody; and  
 (2) a labeled reporter antibody,  
 
 wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a liquid comprising said labeled reporter antibody in the presence of said immobilized capture antibody, and wherein said reagent mixture is stored in said container, and  
 wherein said container is a tube that is capable of being hermetically sealed and wherein said tube comprises an indentation near the open end;  
   (ii) incubating a liquid sample, which may contain said analyte, with said reagent mixture;    (iii) measuring a signal attributable to a complex formed by binding of said immobilized capture antibody and said labeled reporter antibody to said analyte; and    (iv) correlating said measured signal to the presence or amount of said analyte.    
   
   
       25 : The method of  claim 24 , wherein said capture antibody is immobilized on a paramagnetic bead.  
   
   
       26 : The method of  claim 24 , wherein said drying is carried out by lyophilization.  
   
   
       27 : The method of  claim 24 , wherein said signal is an electrochemiluminescent signal.  
   
   
       28 : The method of  claim 24 , wherein said labeled reporter antibody is labeled with a tris bipyridyl ruthenium group.  
   
   
       29 : The method of  claim 24 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       30 : The method of  claim 24 , wherein said tube is a polypropylene tube.  
   
   
       31 : The method of  claim 24 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       32 : The method of  claim 24 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       33 : The method of  claim 24 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.  
   
   
       34 : A reagent mixture, comprising: 
 (1) an immobilized capture antibody; and    (2) a labeled reporter antibody,    wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a mixture of (a) a liquid comprising said labeled reporter antibody and (b) said immobilized capture antibody,    wherein said reagent mixture is contained in a tube which is capable of being hermetically sealed and wherein said tube comprises an indentation near the open end.    
   
   
       35 : The reagent mixture of  claim 34 , wherein said reagent mixture is prepared by a process which comprises: 
 (1) forming a liquid which comprises said labeled reporter antibody;    (2) adding said liquid which comprises said labeled reporter antibody to said tube, wherein said capture antibody is contained in said tube; and    (3) drying said liquid which comprises said labeled reporter antibody, to obtain said reagent mixture.    
   
   
       36 : The reagent mixture of  claim 35 , wherein said drying is lyophilizing.  
   
   
       37 : The reagent mixture of  claim 34 , wherein said liquid is an aqueous solution.  
   
   
       38 : The reagent mixture of  claim 34 , wherein said reagent mixture is an intimate mixture of said labeled reported antibody and said immobilized capture antibody.  
   
   
       39 : The reagent mixture of  claim 34 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       40 : The reagent mixture of  claim 34 , wherein said tube is a polypropylene tube.  
   
   
       41 : The reagent mixture of  claim 34 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       42 : The reagent mixture of  claim 34 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       43 : The reagent mixture of  claim 34 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.  
   
   
       44 : A kit, comprising: 
 (A) a container; and    (B) a reagent mixture comprising: 
 (1) an immobilized capture antibody; and  
 (2) a labeled reporter antibody,  
 wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a liquid comprising said labeled reporter antibody in the presence of said immobilized capture antibody, and wherein said reagent mixture is contained in said container, and  
   wherein said container is a tube which is capable of being hermetically sealed.    
   
   
       45 : The kit of  claim 44 , wherein said capture antibody is immobilized on a paramagnetic bead.  
   
   
       46 : The kit of  claim 44 , wherein said drying is carried out by lyophilization.  
   
   
       47 : The kit of  claim 44 , wherein said labeled reporter antibody is labeled with a tris bipyridyl ruthenium group.  
   
   
       48 : The kit of  claim 44 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       49 : The kit of  claim 44 , wherein said tube is a polypropylene tube.  
   
   
       50 : The kit of  claim 44 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       51 : The kit of  claim 44 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       52 : The kit of  claim 44 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.  
   
   
       53 : A method for detecting or quantifying an analyte, comprising: 
 (i) providing a kit comprising: 
 (A) a container; and  
 (B) a reagent mixture comprising: 
 (1) an immobilized capture antibody; and  
 (2) a labeled reporter antibody,  
 
 wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a liquid comprising said labeled reporter antibody in the presence of said immobilized capture antibody, and wherein said reagent mixture is stored in said container, and  
 wherein said container is a tube that is capable of being hermetically sealed;  
   (ii) incubating a liquid sample, which may contain said analyte, with said reagent mixture;    (iii) measuring a signal attributable to a complex formed by binding of said immobilized capture antibody and said labeled reporter antibody to said analyte; and    (iv) correlating said measured signal to the presence or amount of said analyte.    
   
   
       54 : The method of  claim 53 , wherein said capture antibody is immobilized on a paramagnetic bead.  
   
   
       55 : The method of  claim 53 , wherein said drying is carried out by lyophilization.  
   
   
       56 : The method of  claim 53 , wherein said signal is an electrochemiluminescent signal.  
   
   
       57 : The method of  claim 53 , wherein said labeled reporter antibody is labeled with a tris bipyridyl ruthenium group.  
   
   
       58 : The method of  claim 53 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       59 : The method of  claim 53 , wherein said tube is a polypropylene tube.  
   
   
       60 : The method of  claim 53 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       61 : The method of  claim 53 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       62 : The method of  claim 53 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.  
   
   
       63 : A reagent mixture, comprising: 
 (1) an immobilized capture antibody; and    (2) a labeled reporter antibody,    wherein said immobilized capture antibody and said labeled reporter antibody bind specifically to a same analyte, and wherein said reagent mixture has been prepared by drying a mixture of (a) a liquid comprising said labeled reporter antibody and (b) said immobilized capture antibody,    wherein said reagent mixture is contained in a tube which is capable of being hermetically sealed.    
   
   
       64 : The reagent mixture of  claim 63 , wherein said reagent mixture is prepared by a process which comprises: 
 (1) forming a liquid which comprises said labeled reporter antibody;    (2) adding said liquid which comprises said labeled reporter antibody to said tube, wherein said capture antibody is contained in said tube; and    (3) drying said liquid which comprises said labeled reporter antibody, to obtain said reagent mixture.    
   
   
       65 : The reagent mixture of  claim 64 , wherein said drying is lyophilizing.  
   
   
       66 : The reagent mixture of  claim 63 , wherein said liquid is an aqueous solution.  
   
   
       67 : The reagent mixture of  claim 63 , wherein said reagent mixture is an intimate mixture of said labeled reported antibody and said immobilized capture antibody.  
   
   
       68 : The reagent mixture of  claim 63 , wherein said tube has dimensions of about 12 mm×about 75 mm.  
   
   
       69 : The reagent mixture of  claim 63 , wherein said tube is a polypropylene tube.  
   
   
       70 : The reagent mixture of  claim 63 , wherein said immobilized capture antibody is specific for a  Clostridium botulinum  A neurotoxin antigen.  
   
   
       71 : The reagent mixture of  claim 63 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, a protein, a hormone, a DNA, a RNA, a drug, an antibiotic, or a nerve toxin.  
   
   
       72 : The reagent mixture of  claim 63 , wherein said immobilized capture antibody is specific for a bacterial toxin, a virus, a bacteria, or a nerve toxin.