Immunoassay methods and reagents for decreasing nonspecific binding
First Claim
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1. A method for measuring Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) in a test sample, the method comprising:
- (a) contacting the test sample with a first reagent, wherein the first reagent consists of a solid phase coated with an anti-PIVKA-II antibody, a buffer, a detergent, and bovine serum albumin, wherein the anti-PIVKA-II antibody binds to an epitope on PIVKA-II to form an anti-PIVKA-II antibody-PIVKA-II antigen complex in the presence of the buffer, the detergent, and the bovine serum albumin;
(b) contacting the anti-PIVKA-II antibody-PIVKA-II antigen complex with a second reagent, wherein the second reagent comprises a detectably-labeled anti-prothrombin antibody, 2-(N-morpholino)ethanesulfonic acid (MES) buffer with a bovine protein stabilizer, an antimicrobial agent, and 30 mM-200 mM magnesium chloride (MgCl2), wherein the anti-prothrombin antibody is MCA1-8, binds to an epitope on PIVKA-II, and forms an anti-PIVKA-II antibody-PIVKA-II antigen-anti-prothrombin antibody complex; and
(c) determining the PIVKA-II concentration in the test sample based on the signal generated by the detectable label in the anti-PIVKA-II antibody-PIVKA-II antigen-anti-prothrombin antibody complex formed in (b).
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Abstract
Methods and kits for reducing non-specific binding in an immunoassay for PIVKA-II in a test sample are described, in which the test sample is reacted with an anti-prothrombin antibody in the presence of one or more of the following additives: skim milk, saponin, CaCl2, MgCl2, and a sulfobetaine zwitterionic detergent.
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Citations
2 Claims
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1. A method for measuring Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) in a test sample, the method comprising:
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(a) contacting the test sample with a first reagent, wherein the first reagent consists of a solid phase coated with an anti-PIVKA-II antibody, a buffer, a detergent, and bovine serum albumin, wherein the anti-PIVKA-II antibody binds to an epitope on PIVKA-II to form an anti-PIVKA-II antibody-PIVKA-II antigen complex in the presence of the buffer, the detergent, and the bovine serum albumin; (b) contacting the anti-PIVKA-II antibody-PIVKA-II antigen complex with a second reagent, wherein the second reagent comprises a detectably-labeled anti-prothrombin antibody, 2-(N-morpholino)ethanesulfonic acid (MES) buffer with a bovine protein stabilizer, an antimicrobial agent, and 30 mM-200 mM magnesium chloride (MgCl2), wherein the anti-prothrombin antibody is MCA1-8, binds to an epitope on PIVKA-II, and forms an anti-PIVKA-II antibody-PIVKA-II antigen-anti-prothrombin antibody complex; and (c) determining the PIVKA-II concentration in the test sample based on the signal generated by the detectable label in the anti-PIVKA-II antibody-PIVKA-II antigen-anti-prothrombin antibody complex formed in (b). - View Dependent Claims (2)
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Current AssigneeAbbott Japan LLC (Abbott Laboratories)
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Original AssigneeAbbott Japan Co., Ltd. (Abbott Laboratories)
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InventorsYoshimura, Toru, Yoshida, Eisaku, Chiba, Ryotaro
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Primary Examiner(s)Grossman, Andrea S
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Application NumberUS13/472,683Publication NumberTime in Patent Office2,351 DaysField of SearchUS Class CurrentCPC Class CodesG01N 33/86 involving blood coagulating...
