Method for serological typing using type-specific antigens

Abstract

The invention concerns a method for typing antibodies in a sample liquid by means of type-specific antigens and in particular a method for typing antibodies to the hepatitis C virus and peptide antigens suitable for this.

1 . Method for typing antibodies in a sample liquid, 
 wherein 
 (a) a first aliquot of the sample liquid is contacted with a first immobilized antigen which is specific for a first type of the antibodies to be examined or with a first mixture of immobilized antigens each of which is specific for a first type of antibodies to be examined under conditions in which the antigen or antigen mixture can react with the antibodies and in which the amount of antibody in the sample liquid does not exceed the capacity of the immobilized antigen or antigen mixture,  
 (b) the sample liquid from step (a) is contacted with a second immobilized antigen which is specific for a second type of antibodies to be examined or with a mixture of immobilized antigens each of which is specific for a second type of antibodies to be examined under conditions as in step (a), the second antigen or antigen mixture being spatially separate from the first antigen or antigen mixture used in step (a),  
 (c) the measures according to step (b) are optionally repeated with one or several further antigens or antigen mixtures which are specific for one or several further types of antibodies to be examined, the further antigens or antigen mixtures each being spatially separate from the antigens or antigen mixtures used in previous steps,  
 (d) a second aliquot of the sample liquid is optionally contacted with several immobilized antigens or antigen mixtures according to steps (a) to (c) in which the sequence of antigens or antigen mixtures is, however, different,  
 (e) the respective immunological reactivity of the immobilized antigens or antigen mixtures with the sample liquid is determined qualitatively or/and quantitatively and  
 (f) a typing of the antibodies present in the sample liquid is carried out based on the reactivity determination.  
   
     
     
         2 . Method as claimed in  claim 1 , 
 wherein 
 peptides are used as immobilized antigens.  
   
     
     
         3 . Method as claimed in  claim 1  or  2 , 
 wherein 
 the antigens are immobilized on microtitre plates.  
 
 
     
     
         4 . Method as claimed in one of the  claims 1  to  3 , 
 wherein 
 the antigens carry a solid phase binding group via which they are coupled to a reactive solid phase by means of an affinity interaction.  
 
 
     
     
         5 . Method as claimed in  claim 4 , 
 wherein 
 the solid phase binding group is selected from biotin and biotin derivatives and the solid phase is coated with streptavidin or avidin.  
   
     
     
         6 . Method as claimed in one of the  claims 1  to  3 , 
 wherein 
 the antigens are covalently linked to a solid phase.  
 
 
     
     
         7 . Method as claimed in one of the  claims 1  to  3 , 
 wherein 
 the antigens are present conjugated to a carrier which is adsorptively coupled to a solid phase.  
 
 
     
     
         8 . Method as claimed in one of the  claims 1  to  7 , 
 wherein 
 a typing of antibodies which are directed towards one or several pathogens is carried out.  
 
 
     
     
         9 . Method as claimed in  claim 8 , 
 wherein 
 antiviral antibodies are typed.  
   
     
     
         10 . Method as claimed in  claim 9 , 
 wherein 
 antibodies to hepatitis C virus are typed.  
   
     
     
         11 . Peptide comprising at least one immunologically active region from the hepatitis C virus selected from 
 (a) the amino acids 384-414,    (b) the amino acids 1738-1759,    (c) the amino acids 2217-2236,    (d) the amino acids 2402-2419,    (e) the amino acids 2345-2357    and partial sequences thereof having a length of at least 6 amino acids.    
     
     
         12 . Peptide as claimed in  claim 11 , 
 wherein 
 the immunologically active region is selected from (a) the amino acid sequences shown in SEQ ID NO. 1-10, (b) amino acid sequences which have a homology of at least 90% to one of the sequences from (a), or (c) partial sequences of sequences from (a) or (b) having a length of at least 6 amino acids.  
   
     
     
         13 . Peptide as claimed in  claim 11 , 
 wherein 
 the immunologically active region is selected from (a) the amino acid sequences shown in SEQ ID NO. 11-16, (b) amino acid sequences which have a homology of at least 90% to one of the sequences from (a), or (c) partial sequences of sequences from (a) or (b) with a length of at least 6 amino acids.  
   
     
     
         14 . Peptide as claimed in  claim 11 , 
 wherein 
 the immunologically active region is selected from (a) the amino acid sequences shown in SEQ ID NO. 17-22, (b) amino acid sequences which have a homology of at least 90 t to one of the sequences from (a), or (c) partial sequences of sequences from (a) or (b) with a length of at least 6 amino acids.  
   
     
     
         15 . Peptide as claimed in  claim 11 , 
 wherein 
 the immunologically active region is selected from (a) the amino acid sequences shown in SEQ ID NO. 23-24, (b) amino acid sequences which have a homology of at least 90% to one of the sequences from (a), or (c) partial sequences of sequences from (a) or (b) with a length of at least 6 amino acids.  
   
     
     
         16 . Peptide as claimed in  claim 11 , 
 wherein 
 the immunologically active region is selected from (a) the amino acid sequences shown in SEQ ID NO. 25-30, (b) amino acid sequences which have a homology of at least 90% to one of the sequences from (a), or (c) partial sequences of sequences from (a) or (b) with a length of at least 6 amino acids.  
   
     
     
         17 . Peptide as claimed in one of the  claims 11  to  16 , 
 wherein 
 the immunologically active region has a length of 30 amino acids at most.  
 
 
     
     
         18 . Peptide as claimed in one of the  claims 1  to  17 , 
 wherein 
 the immunologically active region has a length of 9 to 20 amino acids.  
 
 
     
     
         19 . Peptide as claimed in one of the  claims 1  to  18 , 
 wherein 
 it additionally comprises an immunologically inactive spacer region.  
 
 
     
     
         20 . Peptide as claimed in one of the  claims 1  to  19 , 
 wherein 
 it carries at least one solid phase binding group.  
 
 
     
     
         21 . Peptide as claimed in  claim 20 , 
 wherein 
 the solid phase binding group is selected from biotin and biotin derivatives.  
   
     
     
         22 . Peptide as claimed in one of the  claims 1  to  19 , 
 wherein 
 it carries at least one marker group.  
 
 
     
     
         23 . Peptide as claimed in  claim 22 , 
 wherein 
 the marker group is selected from luminescent metal complexes and fluorescent dyes.  
   
     
     
         24 . Use of peptides as claimed in one of the  claims 11  to  23  in a method for the determination of antibodies to the hepatitis C virus.  
     
     
         25 . Use as claimed in  claim 24  as antigens in a diagnostic method for the detection of a HCV infection.  
     
     
         26 . Use as claimed in  claim 24  in a double antigen bridge test.  
     
     
         27 . Use as claimed in  claim 24  in a method for typing antibodies to HCV.  
     
     
         28 . Use as claimed in  claim 27 , 
 wherein 
 the typing method is carried out as claimed in one of the  claims 1  to  7 .