Method for Measuring Glycoprotein, Method for Examining Liver Desease, Reagent for Quantitative Determination of Glycoprotein and Glycan-Marker Glycoprotein as an Index for Clinical Conditions of Liver Disease

Abstract

An object of the present invention is to provide a method for measuring a glycan-marker glycoprotein, by which liver disease can be detected with higher accuracy than is possible with conventional methods. Also, an object of the present invention is to provide a method for examining liver disease, by which liver disease can be detected with higher accuracy than is possible with conventional methods. Also, an object of the present invention is to provide a reagent for quantitative determination of a glycoprotein, which is used for the above measurement methods. Furthermore, an object of the present invention is to provide a glycan-marker glycoprotein as an index for clinical conditions of liver disease, which is capable of identifying the clinical conditions of liver disease depending on the progress of liver disease. The method for measuring a glycoprotein is characterized in that: the glycoprotein is at least one glycoprotein selected from alpha-1-acid glycoprotein (AGP) and Mac-2-binding protein (M2BP) contained in a sample collected from a subject; when the glycoprotein is AGP, AGP binding to a first lectin selected from AOL and MAL is measured; and when the glycoprotein is M2BP, M2BP binding to a second lectin selected from WFA, BPL, AAL, RCA120, and TJAII is measured.

1 - 30 . (canceled) 
     
     
         31 . A reagent for measuring a glycan structure changed Mac-2-binding protein in a Mac-2-binding protein (M2BP) contained in a blood sample collected from a subject, comprising:
 a Wisteria floribunda lectin (WFA); and   an anti-M2BP antibody.   
     
     
         32 . The reagent according to  claim 31 , wherein the WFA selectively binds to the glycan structure changed Mac-2-binding protein in the blood sample and the anti-M2BP antibody binds to the glycan structure changed Mac-2-binding protein bound to the WFA. 
     
     
         33 . The reagent according to  claim 31 , wherein the WFA is immobilized to a particle. 
     
     
         34 . The reagent according to  claim 33 , wherein the particle is a magnetic particle. 
     
     
         35 . The reagent according to  claim 31 , wherein the WFA is immobilized to a plate substrate. 
     
     
         36 . The reagent according to  claim 33 , wherein the WFA is a biotinylated WFA, and the particle is a streptavidin or an avidin immobilized particle. 
     
     
         37 . The reagent according to  claim 31 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody. 
     
     
         38 . The reagent according to  claim 37 , wherein a label of the labeled anti-M2BP antibody is alkaline phosphatase. 
     
     
         39 . The reagent according to  claim 37 , wherein a label of the labeled anti-M2BP antibody is deglycosylated alkaline phosphatase. 
     
     
         40 . The reagent according to  claim 31 , wherein the anti-M2BP antibody is a deglycosylated anti-M2BP antibody. 
     
     
         41 . The reagent according to  claim 31 , further comprising a Datura stramonium lectin (DSA). 
     
     
         42 . A reagent for measuring a glycan structure changed Mac-2-binding protein in a Mac-2-binding protein (M2BP) contained in a blood sample collected from a subject, comprising:
 a Wisteria floribunda lectin (WFA) is immobilized a magnetic particle; and   an anti-M2BP antibody labeled by alkaline phosphatase.   
     
     
         43 . A method for measuring a glycan structure changed Mac-2-binding protein (M2BP) contained in a blood sample collected from a subject, comprising:
 providing a reagent comprising a Wisteria floribunda lectin (WFA) and an anti-M2BP antibody;   contacting the blood sample with the WFA;   binding the anti-M2BP antibody to the glycan structure changed M2BP bound to the WFA; and   measuring an amount of the anti-M2BP antibody bound to the glycan structure changed M2BP.   
     
     
         44 . The method according to  claim 43 , wherein the WFA is immobilized to a particle. 
     
     
         45 . The method according to  claim 44 , wherein the particle is a magnetic particle. 
     
     
         46 . The method according to  claim 43 , wherein the WFA is immobilized to a plate substrate. 
     
     
         47 . The method according to  claim 44 , wherein the WFA is a biotinylated WFA, and the particle is a streptavidin or an avidin immobilized particle. 
     
     
         48 . The method according to  claim 43 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody, and the measuring is performed by measuring an amount of a label of the labeled anti-M2BP antibody bound to the glycan structure changed M2BP. 
     
     
         49 . The method according to  claim 48 , wherein the label of the labeled anti-M2BP antibody is alkaline phosphatase. 
     
     
         50 . The method according to  claim 48 , wherein the label of the labeled anti-M2BP antibody is deglycosylated alkaline phosphatase, and the anti-M2BP antibody is a deglycosylated anti-M2BP antibody.