HCV antigen-antibody combination assay and methods and compositions for use therein

• US 9,841,427 B2
• Filed: 09/11/2015
• Issued: 12/12/2017
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Dawson, George J., Desai, Suresh M., Gutierrez, Robin A., Muerhoff, A. Scott, Prostko, John, Bogdan, M. Felicia, Marohnic, Christopher C.
• Primary Examiner(s): LI, BAO Q
• Application Number:
  US14/851,471
• Publication Number:
  US 20160123983A1
• Time in Patent Office: 823 Days
• CPC Class Codes: G01N 2333/186 Hepatitis C; Hepatitis NANB
• CPC Class Codes: G01N 2333/914 Hydrolases (3)
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 2469/20 Detection of antibodies in sample from host which are directed against antigens from microorganisms
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: G01N 33/5767 non-A, non-B hepatitis

Abstract

The present invention generally relates to combination immunoassays, reagents and kits for simultaneous detection of HCV antigens and anti-HCV antibodies in a test sample.

Claims

1. A method for the combined detection of an HCV antigen and an HCV antibody in a test sample, the method comprising:

a) providing the following components;

i. a solid phase comprising a tagged anti-HCV core monoclonal antibody and a first tagged HCV capture antigen bound thereto, wherein the anti-HCV core monoclonal antibody binds an HCV core antigen in the test sample and the first HCV capture antigen binds a first anti-HCV antibody in the test sample, and wherein the first HCV capture antigen consisting of the amino acid sequence of SEQ ID NO;

97, 98, 99, 100, 102, 103, 104, 105, or 106; and

ii. a first detectably labeled HCV detection antigen for binding to the first anti-HCV antibody captured by the first HCV capture antigen;

b) incubating the components of step (a) under conditions whereby;

i. the tagged anti-HCV core monoclonal antibody specifically binds to the HCV core antigen present in the test sample to produce an anti-HCV core monoclonal antibody-HCV core antigen complex captured on the solid phase;

ii. the first tagged HCV capture antigen specifically binds to the first anti-HCV antibody present in the test sample to produce a first HCV capture antigen-first anti-HCV antibody complex captured on the solid phase; and

iii. the first detectably labeled HCV detection antigen specifically binds to the anti-HCV antibody of the first HCV capture antigen-first anti-HCV antibody complex captured on the solid phase;

c) isolating the solid phase comprising the captured HCV antigen and the first captured anti-HCV antibody from unreacted test sample and reagents;

d) contacting the isolated solid phase with a detectably labeled conjugate antibody that binds to the HCV core antigen of the anti-HCV core monoclonal antibody-HCV core antigen complex; and

e) detecting;

i. a first signal generated from the detectably labeled conjugate antibody, wherein presence of the first signal indicates presence of the HCV core antigen in the test sample; and

ii. a second signal generated from the first detectably labeled HCV detection antigen, wherein presence of the second signal indicates presence of the first anti-HCV antibody in the test sample.

2. The method of claim 1, wherein:

step (a) further comprises;

iii. a second tagged HCV capture antigen bound to the solid phase that is distinct from the first tagged HCV capture antigen, and binds to a second anti-HCV antibody in the test sample, wherein the second HCV capture antigen comprises domains I, II, and III of HCV NS3 helicase;

iv. a second detectably labeled HCV detection antigen for binding to the second anti-HCV antibody, wherein the second HCV detection antigen comprises domains I, II, and III of HCV NS3 helicase;

step (b) further comprises incubating the components of step (a) under conditions whereby;

iv. the second tagged HCV capture antigen specifically binds to the second anti-HCV antibody present in the test sample to produce a second HCV capture antigen-second anti-HCV antibody complex captured on the solid phase;

v. the second detectably labeled HCV detection antigen specifically binds to the second anti-HCV antibody in the second HCV capture antigen-second anti-HCV antibody complex captured on the solid phase; and

step (e) further comprises;

iii. detecting a third signal generated from the second detectably labeled HCV detection antigen, wherein presence of the third signal indicates presence of the second anti-HCV antibody in the test sample.

3. The method of claim 2, wherein:

step (a) further comprises;

v. a third tagged HCV capture antigen bound to the solid phase that is distinct from the first and the second tagged HCV capture antigens and binds to a third anti-HCV antibody present in the test sample, wherein the third HCV capture antigen comprises at least a portion of domain I of HCV NS3 helicase;

vi. a third detectably labeled HCV detection antigen for binding to the third anti-HCV antibody wherein the third HCV detection antigen comprises at least a portion of domain I of HCV NS3 helicase;

step (b) further comprises incubating the components of step (a) under conditions whereby;

vi. the third tagged HCV capture antigen specifically binds to the third anti-HCV antibody present in the test sample to produce a third HCV capture antigen-third anti-HCV antibody complex captured on the solid phase; and

vii. the third detectably labeled HCV detection antigen specifically binds to the third anti- HCV antibody in the third HCV capture antigen-third anti-HCV antibody complex captured on the solid phase; and

step (e) further comprises;

iv. detecting a fourth signal generated from the third detectably labeled HCV detection antigen, wherein presence of the fourth signal indicates presence of the third anti-HCV antibody in the test sample.

4. The method of claim 1, wherein the solid phase comprises avidin or streptavidin, and the first, second, and third HCV capture antigens are tagged with biotin.