Reagents For HCV Antigen-Antibody Combination Assays

Patent metadata

Field	Value
Publication number	US-2016109447-A1
Application number	US-201514982015-A
Country	US
Kind code	A1
Filing date	Dec 29, 2015
Priority date	May 20, 2009
Publication date	Apr 21, 2016
Grant date	—

Abstract

The present invention is directed to combination immunoassays, reagents and kits for simultaneous detection of HCV antigens and anti-HCV antibodies in a sample. The combination immunoassays of the present invention employ a non-ionic detergent that effectively exposes or releases the HCV core antigen from virions in a sample without interfering with the performance of other reagents such as the capture of anti-HCV antibodies by recombinant HCV antigens.

First claim

What is claimed is:

1 . An immunoassay for simultaneously detecting HCV antigens and antibodies in a sample, comprising:
providing a non-ionic detergent comprising an N-alkyl-N,N-dimethyl-amine oxide, a first pair of a capture antigen and a detection antigen, a first pair of a capture antibody and a conjugate antibody, wherein said capture antigen and said detection antigen both comprise a first peptide fragment of a first HCV protein, said capture antibody and said conjugate antibody specifically bind to a second HCV protein, and said detection antigen and said conjugate antibody comprise one and same signal generating means; contacting said sample in the presence of said detergent with said capture antigen, said detection antigen, said capture antibody and said conjugate antibody, to form a sandwich complex between said capture antigen, said detection antigen, and an anti-HCV antibody present in said sample, and a complex between said capture antibody, said conjugate antibody, and an HCV antigen present in said sample; and measuring a signal generated from said signal-generating means as a result of the formation of said complexes, thereby simultaneously detecting HCV antigens and antibodies in said sample.

2 . The immunoassay of claim 1 , wherein said N-alkyl-N,N-dimethyl-amine oxide is characterized by the formula, CH 3 —(CH 2 )n-N + —(CH 3 ) 2 O − , wherein n falls in the range of 9 to 13.

3 . The immunoassay of claim 2 , wherein said N-alkyl-N,N-dimethyl-amine oxide is Lauryldimethylamine N-oxide (LDAO).

4 . The immunoassay of claim 1 , wherein said first HCV protein and second HCV protein are independently selected from the group consisting of the core antigen, E1, E2, NS2, NS3, NS4, and NS5.

5 . The immunoassay of claim 1 , wherein said first HCV protein and said second HCV protein are the same, and said capture antibody and said conjugate antibody bind to a region of said second HCV protein outside of said first peptide fragment.

6 . The immunoassay of claim 5 , wherein said first HCV protein and said second HCV protein are both the HCV core antigen.

7 . The immunoassay of claim 1 , wherein a second pair of a capture antigen and a detection antigen is provided, wherein said capture antigen and said detection antigen of the second pair both comprise a second peptide fragment of an HCV protein, wherein said second peptide fragment is different from said first peptide fragment.

8 . The immunoassay of claim 7 , wherein said first peptide fragment and said second peptide fragment are derived from different HCV proteins.

9 . The immunoassay of claim 8 , wherein at least one of said first peptide fragment or said second peptide fragment is a fragment of the HCV core antigen.

10 . The immunoassay of claim 1 , wherein said capture antibody in said first pair comprises two or more antibodies.

11 . The immunoassay of claim 1 , wherein a second pair of a capture antibody and a conjugate antibody is provided, wherein said capture antibody and said conjugate antibody in said second pair specifically bind to said second HCV protein or a different HCV protein.

12 . The immunoassay of claim 1 , wherein said capture antigen and said capture antibody are attached to a solid phase.

13 . A kit for simultaneously detecting HCV antigens and antibodies in a sample, comprising a non-ionic detergent comprising an N-alkyl-N,N-dimethyl-amine oxide, a first pair of a capture antigen and a detection antigen, a first pair of a capture antibody and a conjugate antibody, wherein said capture antigen and said detection antigen comprise a first peptide fragment of a first HCV protein, said capture antibody and said conjugate antibody specifically bind to a second HCV protein, and said detection antigen and said conjugate antibody comprise one and same signal generating means.

14 . The kit of claim 13 , wherein said N-alkyl-N,N-dimethyl-amine oxide is characterized by the formula, CH 3 —(CH 2 )n-N + —(CH 3 ) 2 O − , wherein n falls in the range of 9 to 13.

15 . The kit of claim 14 , wherein said N-alkyl-N,N-dimethyl-amine oxide is Lauryldimethylamine N-oxide (LDAO).

16 . The kit of claim 13 , wherein said first HCV protein and said second HCV protein are independently selected from the group consisting of the core antigen, E1, E2, NS2, NS3, NS4, and NS5.

17 . The kit of claim 13 , wherein said first HCV protein and said second HCV protein are the same, and said capture antibody and said conjugate antibody bind to a region of said second HCV protein outside of said first peptide fragment.

18 . The kit of claim 17 , wherein said first HCV protein and said second HCV protein are both the HCV core antigen.

19 . The kit of claim 13 , further comprising a second pair of a capture antigen and a detection antigen is provided, wherein said capture antigen and said detection antigen of the second pair comprise a second peptide fragment of an HCV protein, wherein said second peptide fragment is different from said first peptide fragment.

20 . The kit of claim 19 , wherein said first peptide fragment and said second peptide fragment are derived from different HCV proteins.

21 . The kit of claim 20 , wherein at least one of said first peptide fragment or said second peptide fragment is a fragment of the HCV core antigen.

22 . The kit of claim 13 , wherein said capture antibody in said first pair comprises two or more antibodies.

23 . The kit of claim 13 , further comprising a second pair of a capture antibody and a conjugate antibody, wherein said capture antibody and said conjugate antibody in said second pair specifically bind to said second HCV protein or a different HCV protein.

24 . The kit of claim 13 , wherein said capture antigen and said capture antibody are attached to a solid phase.

Assignees

• Ortho Clinical Diagnostics Inc

Inventors

• Zheng Jian
• Yang Jianping

Classifications

• G01N2469/10

  Detection of antigens from microorganism in sample from host · CPC title

• G01N2469/20

  Detection of antibodies in sample from host which are directed against antigens from microorganisms · CPC title

• G01N33/56983 Primary

  Viruses · CPC title

• G01N2333/186

  Hepatitis C; Hepatitis NANB · CPC title

• G01N33/5306

  Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding · CPC title