Method of detecting an antibody in a liquid sample

Abstract

The invention relates to a method of evaluating the immunological status of a subject comprising the steps of 1 ) determining the content of an antibody in a liquid sample from the subject using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the presence of other constituents of the sample to obtain a measurement 1, 2 ) determining the content of an antibody in the liquid sample using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the absence of other constituents of the sample to obtain a measurement 2, and 3 ) interrelating measurements 1 and 2 to express the interference and using the interference as a parameter for evaluating the immunological status of the subject.

1 - 37 . (canceled)  
   
   
       38 . A method of evaluating and/or predicting the effect of a Specific Allergy Vaccination treatment comprising the steps of 
 (x′) determining the content of an antibody in a liquid sample using the following method: 
 (o′) providing a mixture of a liquid phase and a two-component solid phase complex composed of (i) the antibody of the sample, and (ii) a reactant antibody directed against the sample antibody, the reactant antibody being bound to a solid particle,  
 (p′) separating the two-component solid phase complex from the liquid phase,  
 (q′) washing the separated two-component solid phase complex to remove non-complex bound compounds,  
 (r′) adding to the washed two-component solid phase complex a solution of (iii) a ligand in the form of an antigen, an antibody or a hapten, which is optionally labelled, to form a three-component solid phase complex,  
 (s′) optionally adding to the three-component solid phase complex a solution of (iv) a label compound to form a four-component solid phase complex,  
 (t′) separating the three- or four-component solid phase complex obtained in step (r′) or (s′), respectively, from the solution,  
 (u′) washing the separated three- or four-component solid phase complex to remove non-complex bound compounds,  
 (v′) performing a detection/measurement of the washed labelled three- or four-component complex,  
   (y′) determining the content of the said antibody using the following assay: 
 (ya′) providing a mixture of a liquid phase and a three-component solid phase complex composed of (i) the antibody of the sample, (ii) a reactant antibody directed against the sample antibody, the reactant antibody being bound to a solid particle, (iii) a ligand in the form of an antigen, an antibody or a hapten, which is labelled or bound to (iv) a label compound, to form a multi-component solid phase complex,  
 (yb′) separating the multi-component solid phase complex from the liquid phase,  
 (yc′) washing the separated multi-component solid phase to remove non-complex bound compounds,  
 (yd′) performing a detection/measurement of the washed labelled multi-component complex.  
   (z′) comparing the measurements obtained in step (x′) and step (y′) and using the comparison to evaluate and/or predict the effect of the Specific Allergy Vaccination treatment.    
   
   
       39 . The method according to  claim 38 , 
 wherein in step (r′) and in step (ya′) the ligand is bound to biotin or a functional derivative thereof, and    wherein step (s′) is mandatory and wherein in step (s′) and in step (ya′) the label compound is a chemiluminescent compound covalently bound to avidin, streptavidin or a functional derivative thereof, and    wherein in step (v′) and in step (yd′) the detection/measurement is carried out by initiating a chemiluminescent reaction in the washed four-component solid phase complex and detecting/measuring the resulting chemiluminescence, if any.    
   
   
       40 . The method according to  claim 39 , 
 wherein in step (o′) and in step (ya′) the solid particle is a solid paramagnetic particle, and    wherein in step (p′), in step (t′) and in step (yb′) the separation of the solid phase complex from the liquid phase is performed magnetically.    
   
   
       41 . The method according to  claim 38 , wherein step (ya′) is carried out by mixing components (i) and (ii), then adding component (iii), and finally adding component (iv), if added.  
   
   
       42 . The method according to  claim 38 , wherein step (ya′) is carried out by mixing components (i), (ii) and (iii), and then adding component (iv), if added.  
   
   
       43 . The method according to  claim 38 , wherein the comparison of step (z′) is carried out by calculating the ratio of the measurements obtained in the two said steps.  
   
   
       44 . The method according to  claim 38 , wherein the comparison of step (z′) is carried out at a number of points in time at the start of and during the treatment period, and that any temporal change, which may be observed, is used as a basis for evaluating and/or predicting the effect of the treatment.  
   
   
       45 . The method according to  claim 38 , wherein the label compound is selected from the group consisting of a luminescent label, a chemiluminescent label, an enzyme label, a radioactivity label, a fluorescent label and an absorbance label.  
   
   
       46 . The method according to  claim 38 , wherein the labeled ligand is labeled by a radioactive atom.  
   
   
       47 . The method according to  claim 38 , wherein the separation of the solid phase complex from the liquid phase is carried out by a member selected from the group consisting of magnetic separation, filtration, sedimentation, centrifugation, chromatography and column chromatography.  
   
   
       48 . The method according to  claim 38 , wherein component (iii) of step (r′) and component (iv) of step (s′), respectively, are added in one operation.  
   
   
       49 . The method according to  claim 39 , wherein the chemiluminescent compound is an acridinium compound.  
   
   
       50 . The method according to  claim 38 , wherein the three-component solid phase complex formed in step (r′) prior to subjecting it to step (s′) is washed to remove non-complex bound compounds.  
   
   
       51 . A method of detecting and/or quantifying an antibody in a liquid sample comprising the steps of: 
 (o′) providing a mixture of a liquid phase and a two-component solid phase complex composed of (i) the antibody of the sample, and (ii) a reactant antibody directed against the sample antibody, the reactant antibody being bound to a solid particle;    (p′) separating the two-component solid phase complex from the liquid phase;    (q′) washing the separated two-component solid phase complex to remove non-complex bound compounds;    (r′) adding to the washed two-component solid phase complex a solution of    (iii) a ligand in the form of an antigen, an antibody or a hapten, which is optionally labeled, to form a three-compound solid phase complex;    (s′) optionally adding to the three-component solid phase complex a solution of (iv) a label compound to form a four-component solid phase complex;    (t′) separating the three- or four-component solid phase complex obtained in step (r′) or (s′), respectively, from the solution;    (u′) washing the separated multi-component solid phase complex to remove non-complex bound compounds; and    (v′) performing a detection/measurement of the washed labeled multi-component complex.    
   
   
       52 . The method according to  claim 51 , wherein in step (r′) the ligand is bound to biotin or a functional derivative thereof, and 
 wherein step (s′) is mandatory and wherein in step (s′) the label compound is a chemiluminescent compound covalently bound to avidin, streptavidin or a functional derivative thereof, and    wherein in step (v′) the detection/measurement is carried out by initiating a chemiluminescent reaction in the washed four-component solid phase complex and detecting/measuring the resulting chemiluminescence, if any.    
   
   
       53 . The method according to  claim 52 , wherein in step (o′) the solid particle is a solid paramagnetic particle, and 
 wherein in step (p′) and in step (t′) the separation of the solid phase complex from the liquid phase is performed magnetically.    
   
   
       54 . The method according to  claim 52 , wherein the chemiluminescent compound is an acridinium compound.  
   
   
       55 . The method according to  claim 51 , wherein component (iii) of step (r′), and component (iv) of step (s′), respectively, are added in one operation.  
   
   
       56 . The method according to  claim 51 , wherein the three-component solid phase complex formed in step (r′) prior to subjecting it to step(s′) is washed to remove non-complex bound compounds.