Method for measuring glycoprotein, method for examining liver disease, 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 method for detecting cirrhosis, comprising:
 mixing a blood sample collected from a patient with liver disease, a  Wisteria floribunda  lectin (WFA) and an anti-Mac-2-binding protein antibody;   assaying Mac-2-binding protein (M2BP) binding to both the WFA and the anti-M2BP antibody; and   determining whether or not the liver disease is cirrhosis based on the assayed value of the M2BP.   
     
     
         32 . The method according to  claim 31 , wherein the blood sample is a serum. 
     
     
         33 . The method according to  claim 31 , wherein the mixing is performed by mixing the blood sample and the WFA, and mixing the M2BP antibody and a mixture of the blood sample and the WFA. 
     
     
         34 . The method according to  claim 31 , wherein the WFA is immobilized on a particle. 
     
     
         35 . The method according to  claim 34 , wherein the particle is a magnetic particle. 
     
     
         36 . The method according to  claim 35 , wherein the WFA is a biotinylated WFA, and the magnetic particle is a streptavidin or an avidin immobilized magnetic particle. 
     
     
         37 . The method according to  claim 31 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody, and a label of the labeled anti-M2BP antibody is a fluorescent substance or an enzyme. 
     
     
         38 . The method according to  claim 37 , wherein the fluorescent substance is selected from a fluorescein isothiocyanate, a green fluorescent protein and a luciferin. 
     
     
         39 . The method according to  claim 37 , wherein the enzyme is selected from an alkaline phosphatase, a peroxidase, a glucose oxidase, a tyrosinase and an acid phosphatase. 
     
     
         40 . The method according to  claim 37 , wherein the enzyme is the alkaline phosphatase, which is assayed by using a luminescent substrate or a chromogenic substrate. 
     
     
         41 . The method according to  claim 31 , wherein the WFA selectively binds to glycoproteins having a specific glycan structure in the blood sample, the anti-M2BP antibody binds to a core protein of Mac-2-binding protein, and a glycoprotein binding both the WFA and the anti-M2BP antibody is the glycan structure changed Mac-2-binding protein. 
     
     
         42 . A method for distinguishing cirrhosis and hepatitis, comprising:
 providing a  Wisteria floribunda  lectin (WFA) and an anti-Mac-2-binding protein antibody;   assaying Mac-2-binding protein (M2BP) which is contained in a blood sample collected from a patient with liver disease and binds to both the WFA and the anti-M2BP antibody; and   determining whether the liver disease is cirrhosis or hepatitis based on the assayed value of the M2BP.   
     
     
         43 . The method according to  claim 42 , wherein the blood sample is a serum sample. 
     
     
         44 . The method according to  claim 42 , wherein the WFA is immobilized on a particle. 
     
     
         45 . The method according to  claim 44 , wherein the particle is a magnetic particle. 
     
     
         46 . The method according to  claim 42 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody, and a label of the labeled anti-M2BP antibody is a fluorescent substance or an enzyme. 
     
     
         47 . The method according to  claim 46 , wherein the enzyme is the alkaline phosphatase, which is assayed by using a luminescent substrate or a chromogenic substrate. 
     
     
         48 . The method according to  claim 42 , wherein the WFA selectively binds to glycoproteins having a specific glycan structure in the blood sample, the anti-M2BP antibody binds to a core protein of Mac-2-binding protein, and a glycoprotein sandwiched by the WFA and the anti-M2BP antibody is the glycan structure changed Mac-2-binding protein. 
     
     
         49 . A method for detecting cirrhosis, comprising:
 providing a  Wisteria floribunda  lectin (WFA) and an anti-Mac-2-binding protein antibody;   assaying Mac-2-binding protein (M2BP) which is contained in a blood sample collected from a patient with liver disease and binds to both the WFA and the anti-M2BP antibody; and   determining whether or not the liver disease is cirrhosis based on the assayed value of the M2BP.   
     
     
         50 . The method according to  claim 49 , wherein the blood sample is a serum. 
     
     
         51 . The method according to  claim 49 , wherein the WFA is immobilized on a particle. 
     
     
         52 . The method according to  claim 51 , wherein the particle is a magnetic particle. 
     
     
         53 . The method according to  claim 49 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody, and a label of the labeled anti-M2BP antibody is a fluorescent substance or an enzyme. 
     
     
         54 . The method according to  claim 49 , wherein the enzyme is the alkaline phosphatase, which is assayed by using a luminescent substrate or a chromogenic substrate. 
     
     
         55 . A method for detecting a risk of recurrence of hepatic cell carcinoma, comprising:
 providing a  Wisteria floribunda  lectin (WFA) and an anti-Mac-2-binding protein antibody;   assaying Mac-2-binding protein (M2BP) which is contained in a blood sample collected from a patient with hepatic cell carcinoma after operation and binds to both the WFA and the anti-M2BP antibody; and   determining whether the patient has a high possibility of non-recurrence of hepatic cell carcinoma based on the assayed value of the M2BP.   
     
     
         56 . The method according to  claim 55 , wherein the blood sample is a serum. 
     
     
         57 . The method according to  claim 55 , wherein the WFA is immobilized on a particle. 
     
     
         58 . The method according to  claim 57 , wherein the particle is a magnetic particle. 
     
     
         59 . The method according to  claim 55 , wherein the anti-M2BP antibody is a labeled anti-M2BP antibody, and a label of the labeled anti-M2BP antibody is a fluorescent substance or an enzyme.