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CN-122029427-A - Method for measuring GPC3 and kit therefor

CN122029427ACN 122029427 ACN122029427 ACN 122029427ACN-122029427-A

Abstract

Disclosed are a method for measuring GPC3 and an immunoassay kit, wherein the increase in the count value of a negative sample is reduced. The method for measuring GPC3 is a method for measuring GPC3 by an immunoassay, which comprises subjecting GPC3 contained in a liquid sample isolated from a living body to an antigen-antibody reaction with a labeled anti-GPC 3 antibody or antigen-binding fragment thereof labeled with a labeling substance, which comprises bringing a solution containing the labeled anti-GPC 3 antibody or antigen-binding fragment thereof and a polyoxyethylene-polyoxypropylene block copolymer into contact with GPC3 in the sample.

Inventors

  • NISHII TOMONORI
  • KATAGIRI NORIKO
  • Qingliu Keji
  • JIN ZIDUN
  • Ye Daojiu
  • TADOKORO NANAMI

Assignees

  • 富士瑞必欧株式会社

Dates

Publication Date
20260512
Application Date
20241105
Priority Date
20231106

Claims (9)

  1. 1. A method for measuring GPC3, characterized in that, The method is a method for measuring GPC3 by an immunoassay, comprising the step of reacting GPC3 contained in a liquid sample isolated from a living body with a labeled anti-GPC 3 antibody or antigen-binding fragment thereof labeled with a labeling substance, The method comprises contacting a solution comprising the labeled anti-GPC 3 antibody or antigen-binding fragment thereof and a polyoxyethylene-polyoxypropylene block copolymer with GPC3 in the sample.
  2. 2. The method of claim 1, wherein, The immunoassay is a sandwich method.
  3. 3. The method according to claim 1 or 2, wherein, The polyoxyethylene-polyoxypropylene block copolymer has an ethyleneoxy group content of 70% or more and 90% or less.
  4. 4. The method of claim 3, wherein, The polyoxyethylene-polyoxypropylene block copolymer has an ethylene oxide addition mole number of 150-300.
  5. 5. The method according to claim 1 or 2, wherein, The concentration of the polyoxyethylene-polyoxypropylene block copolymer in the solution containing the labeled anti-GPC 3 antibody or antigen-binding fragment thereof and the polyoxyethylene-polyoxypropylene block copolymer is 0.01w/v% to 6.0w/v%.
  6. 6. The method according to claim 1 or 2, wherein, The labeling substance is an enzyme.
  7. 7. A kit for GPC3 measurement, comprising a solution containing an anti-GPC 3 antibody labeled with a labeling substance and a polyoxyethylene-polyoxypropylene block copolymer.
  8. 8. Use of a solution comprising an anti-GPC 3 antibody labeled with a label and a polyoxyethylene-polyoxypropylene block copolymer for GPC3 determination.
  9. 9. Use of a solution comprising an anti-GPC 3 antibody labeled with a label and a polyoxyethylene-polyoxypropylene block copolymer for the manufacture of a kit for GPC3 measurement.

Description

Method for measuring GPC3 and kit therefor Technical Field The present invention relates to a method for measuring GPC3 and a kit therefor. Background Glypican-3 (GPC 3) is a protein expressed in malignant transformation of hepatocytes, and exists in a state immobilized on the cell membrane of hepatocytes. In GPC3, arginine at position 358 and serine at position 359 are cleaved by furin (furin cleavage site), and separated into an N-terminal side subunit of arginine from N-terminus to position 358 and a C-terminal side subunit of serine from position 359 to C-terminus. The N-terminal side subunit and the C-terminal side subunit are bound by 1 intramolecular disulfide bond. Furthermore, GPC3 is a protein that has been confirmed to be specifically expressed in cancers such as hepatocellular carcinoma, and thus a method useful for examining cancer patients targeting GPC3 has been developed. For example, patent document 1 proposes a method for examining a cancer patient by measurement using GPC 3. In addition, patent document 2 proposes an immunoassay method of GPC3 using two different antibodies that bind to different epitopes present in the N-terminal region of GPC 3. Patent document 3 proposes a method for measuring GPC3 by subjecting a sample containing GPC3 to a reduction treatment. Prior art literature Patent literature Patent document 1 International publication No. 2004/038420 Patent document 2 International publication No. 2015/097928 Patent document 3 International publication No. 2022/154119 Disclosure of Invention Technical problem to be solved by the invention The present inventors have found that, when GPC3 is measured by an immunoassay method in a liquid sample isolated from a living body, there is a technical problem that the count value of the liquid sample (negative sample) from a healthy person increases. Accordingly, an object of the present invention is to provide a method for measuring GPC3 and an immunoassay kit, which reduce the increase in the count value of a negative sample. Technical scheme for solving technical problems As a result of intensive studies, the inventors of the present invention have found that, in a method for measuring GPC3 by an immunoassay, by adding a polyoxyethylene-polyoxypropylene block copolymer to a solution containing a labeled antibody, the increase in the count value of a negative sample can be reduced, and completed the present invention. Namely, the present invention provides the following means. (1) A method for measuring GPC3, characterized in that, The method is a method for measuring GPC3 by an immunoassay, comprising the step of reacting GPC3 contained in a liquid sample isolated from a living body with a labeled anti-GPC 3 antibody or antigen-binding fragment thereof labeled with a labeling substance, The method comprises contacting a solution comprising the labeled anti-GPC 3 antibody or antigen-binding fragment thereof and a polyoxyethylene-polyoxypropylene block copolymer with GPC3 in the sample. (2) The method according to (1), wherein the immunoassay is a sandwich method. (3) The method according to (1) or (2), wherein the polyoxyethylene-polyoxypropylene block copolymer has an ethyleneoxy group content of 70% or more and 90% or less. (4) The method according to (3), wherein the polyoxyethylene-polyoxypropylene block copolymer has an addition mole number of ethylene oxide of 150 or more and 300 or less. (5) The method according to any one of (1) to (4), wherein a concentration of the polyoxyethylene-polyoxypropylene block copolymer in the solution containing the labeled anti-GPC 3 antibody or an antigen-binding fragment thereof and the polyoxyethylene-polyoxypropylene block copolymer is 0.01w/v% to 6.0w/v%. (6) The method according to any one of (1) to (5), wherein the labeling substance is an enzyme. (7) A kit for GPC3 measurement, comprising a solution containing an anti-GPC 3 antibody labeled with a labeling substance and a polyoxyethylene-polyoxypropylene block copolymer. (8) Use of a solution comprising an anti-GPC 3 antibody labeled with a label and a polyoxyethylene-polyoxypropylene block copolymer for GPC3 determination. (9) Use of a solution comprising an anti-GPC 3 antibody labeled with a label and a polyoxyethylene-polyoxypropylene block copolymer for the manufacture of a kit for GPC3 measurement. Effects of the invention According to the present invention, a method for measuring GPC3 and a GPC3 measurement kit in which an increase in the count value of a negative sample is suppressed can be provided. Detailed Description The method of the present invention is a method of measuring GPC3 by mixing GPC3 contained in a liquid sample isolated from a living body with a solution containing an anti-GPC 3 antibody labeled with a labeling substance and a polyoxyethylene-polyoxypropylene block copolymer. As shown in the examples, it was found that when GPC3 in a liquid sample isolated from a living body was measured by an immunoassay method, t