Calibration reagent and method

Abstract

A multiplex assay system for detection of a molecule of interest, such as (a) an immunoglobulin or (b) a peptide/protein biomarker present in a biological sample, comprises: —a reaction vessel, a plurality of capturing agents immobilised on a solid phase, a detection molecule, such as (a) an anti-immunoglobulin detection molecule or (b) a detection molecule binding to a peptide/protein biomarker, a calibration reagent, —a reaction buffer medium, characterised in that the calibration reagent comprises at least two different binding molecules, wherein each binding molecule has a capacity to bind specifically to a capturing agent immobilised on the solid phase and a capacity to bind to the detection molecule, and wherein at least two of the binding molecules are present at different concentrations in the calibration reagent.

1 . A multiplex assay system for detection of a molecule of interest, such as (a) an immunoglobulin or (b) a peptide/protein biomarker present in a biological sample,
 comprising:
 a reaction vessel, 
   a plurality of capturing agents immobilised on a solid phase,   a detection molecule, such as (a) an anti-immunoglobulin detection molecule or (b) a detection molecule binding to a peptide/protein biomarker,   a calibration reagent,
 a reaction buffer medium, 
   characterised in that the calibration reagent comprises at least two different binding molecules, wherein each binding molecule has a capacity to bind specifically to a capturing agent immobilised on the solid phase and a capacity to bind to the detection molecule, and wherein at least two of the binding molecules are present at different concentrations in the calibration reagent.   
     
     
         2 . The system of  claim 1  wherein the molecule of interest is (i) an IgE antibody or (ii) an IgG antibody, or (iii) a peptide/protein biomarker for disease. 
     
     
         3 . The system of  claim 1  wherein the detection molecule is (i) an anti-human IgE conjugate, (ii) an anti-human IgG conjugate, or (iii) an antibody specific for a peptide/protein biomarker. 
     
     
         4 . The system of  claim 1  wherein the binding molecules are recombinant antibodies such as chimeric antibodies, native antibodies such as autoantibodies, or peptide/protein biomarkers. 
     
     
         5 . The system of  claim 1  wherein the capturing agents are allergen components, such as native or recombinant allergen components, or disease-related antigens, such as antigenic components related to infectious disease, or antigenic components related to autoimmune disease, or antibodies specific for peptide/protein biomarkers. 
     
     
         6 . The system of  claim 1  wherein the calibration reagent comprises fifteen different mouse-human chimeric IgE antibodies, wherein each chimeric antibody has a capacity to bind specifically to an allergen component selected from the group consisting of Bet v 1, Der p 2, Ole e 1, Gal d 1, Art v 1, Fel d 1, Phi p 1, Amb a 1, Can f 1, Der p 1, Gal d 2, Can f 2, Can f 5, Phi p 5 and Pru p 3. 
     
     
         7 . A calibration reagent comprising at least two different chimeric antibodies, wherein each binding molecule has the capacity to bind specifically to at least one of the allergen components listed in Table 1. 
     
     
         8 . The calibration reagent of  claim 7  comprising at least 5 different mouse-human chimeric IgE antibodies. 
     
     
         9 . The calibration reagent of  claim 7  comprising at least 10 different mouse-human chimeric IgE antibodies. 
     
     
         10 . The calibration reagent of  claim 7  comprising at least 15 different mouse-human chimeric IgE antibodies. 
     
     
         11 . The calibration reagent of  claim 8  wherein each chimeric antibody has the capacity to bind specifically to an allergen component selected from the group consisting of Bet v 1, Der p 2, Ole e 1, Gal d 1, Art v 1, Fel d 1, Phi p 1, Amb a 1, Can f 1, Der p 1, Gal d 2, Can f 2, Can f 5, Phi p 5 and Pru p 3. 
     
     
         12 . The calibration reagent of  claim 11  consisting of fifteen different mouse-human chimeric IgE antibody solutions, wherein each chimeric antibody has the capacity to bind specifically to an allergen component selected from the group consisting of Bet v 1, Der p 2, Ole e 1, Gal d 1, Art v 1, Fel d 1, Phi p 1, Amb a 1, Can f 1, Der p 1, Gal d 2, Can f 2, Can f 5, Phi p 5 and Pru p 3. 
     
     
         13 . A kit comprising a calibration reagent according to  claim 7  and a detection molecule which has a capacity to bind to each antibody in the calibration reagent.