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

• US 20120172247A1
• Filed: 01/13/2012
• Published: 07/05/2012
• Est. Priority Date: 07/14/2009
• Status: Active Grant

• Current Assignee: National Institute of Advanced Industrial Science and Technology (Government of Japan), National Center for Global Health and Medicine, Public University Corporation Nagoya City University, Sysmex Corporation
• Original Assignee: National Institute of Advanced Industrial Science and Technology (Government of Japan), National Center for Global Health and Medicine, Public University Corporation Nagoya City University, Sysmex Corporation
• Inventors: Narimatsu, Hisashi, Ikehara, Yuzuru, Kuno, Atsushi, Tanaka, Yasuhito, Sogabe, Maki, Mizokami, Masashi, Ito, Kiyoaki, Matsubara, Shunsuke, Tsuruno, Chikayuki, Takahama, Youichi, Kagawa, Takashi, Nagai, Shinya
• Granted Patent: US 8,623,608 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: C07K 14/4726 Lectins
• CPC Class Codes: C07K 14/473 alpha-Glycoproteins
• CPC Class Codes: G01N 2333/42 Lectins, e.g. concanavalin, phytohaemagglutinin
• CPC Class Codes: G01N 2333/4724 Lectins
• CPC Class Codes: G01N 2333/4728 alpha-Glycoproteins
• CPC Class Codes: G01N 2400/00 Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
• CPC Class Codes: G01N 2400/02 involving antibodies to sugar part of glycoproteins lectins from plants G01N2333/42, lectins from mammals G01N2333/4724
• CPC Class Codes: G01N 2800/085 Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/57438 of liver, pancreas or kidney
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

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. Disclosed is a method for measuring 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, comprising: measuring AGP binding to a first lectin selected from AOL and MAL, when the glycoprotein is AGP; and measuring M2BP binding to a second lectin selected from WFA, BPL, AAL, RCA120, and TJAII, when the glycoprotein is M2BP.

Claims

1. A method for measuring a glycoprotein comprising:

contacting a sample collected from a subject with a lectin selected from WFA, BPL, AAL, RCA120, and TJAII; and

measuring Mac-2-binding protein (M2BP) binding to the lectin.

2. The method for measuring a glycoprotein according to claim 1, comprising measuring M2BP binding to a lectin for normalization of which reactivity is not substantially altered regardless of glycan structural changes in M2BP and normalizing the measured value of M2BP binding to the lectin using the measured value of M2BP binding to the lectin for normalization.

3. The method for measuring a glycoprotein according to claim 2, wherein the lectin for normalization is DSA.

4. The method for measuring a glycoprotein according to claim 1, wherein the contacting is performed by contacting the sample, the lectin and a labeled anti- M2BP antibody, and forming a complex of the lectin, the M2BP, and the labeled anti- M2BP antibody,

wherein the measuring is performed by measuring the amount of the label of the complex, and

quantitatively determining the M2BP binding to the lectin.

5. The method for measuring a glycoprotein according to claim 4, wherein the complex is formed using magnetic particles to which the lectin is immobilized.

6. The method for measuring a glycoprotein according to claim 4, wherein the complex is formed using the biotinylated lectin and magnetic particles to which streptavidin or avidin are immobilized.

7. The method for measuring a glycoprotein according to claim 4, wherein the label of the labeled anti-M2BP antibody is deglycosylated alkaline phosphatase.

8. The method for measuring a glycoprotein according to claim 4, wherein the anti-M2BP antibody is a deglycosylated anti-M2BP antibody.

9. The method for measuring a glycoprotein according to claim 1, wherein the measuring is performed by measuring M2BP binding to the lectin with the use of a lectin array to which at least the lectin is immobilized and an anti-M2BP antibody.

11. A method for examining liver disease, whereby the stage of fiber formation in the liver is determined based on the measured value of M2BP obtained by the method for measuring M2BP according to claim 1.

12. A method for examining liver disease, whereby whether or not the disease is cirrhosis based on the measured value of M2BP obtained by the method for measuring M2BP according to claim 1.

13. A method for examining liver disease, whereby a risk of recurrence of hepatic cell carcinoma is determined based on the measured value of M2BP obtained by the method for measuring M2BP according to claim 1.

14. A method for determining the therapeutic effect of an anti-viral agent in treatment for liver disease, based on the measured value of M2BP obtained by the method for measuring M2BP according to claim 1.

10. The method for measuring a glycoprotein according to claim 10, wherein:

DSA is further immobilized to the lectin array;

M2BP binding to DSA is measured and then the measured value of M2BP binding to the lectin is normalized using the measured value of M2BP binding to DSA.

26. The method for measuring a glycoprotein according to claim 10, wherein:

DSA is further immobilized to the lectin array;

AGP binding to DSA is measured and then the measured value of AGP binding to the lectin is normalized using the measured value of AGP binding to DSA.

15. A reagent for quantitative determination of a Mac-2-binding protein (M2BP), comprising magnetic particles to which a lectin selected from WFA, BPL, AAL, RCA120, and TJAII is immobilized and a labeled anti-M2BP antibody.

16. A glycan marker as an index for clinical conditions of liver disease, comprising a Mac-2-binding protein (M2BP) having liver-disease-specific glyco-alterations and binding to a lectin selected from WFA, BPL, AAL, RCA120, and TJAII.

17. A method for measuring a glycoprotein comprising:

contacting a sample collected from a subject with a lectin selected from AOL and MAL; and

measuring alpha-1-acid glycoprotein (AGP) binding to the lection.

18. The method for measuring a glycoprotein according to claim 17, comprising measuring AGP binding to a lectin for normalization of which reactivity is not substantially altered regardless of glycan structural changes in AGP and normalizing the measured value of AGP binding to the lectin using the measured value of AGP binding to the lectin for normalization.

19. The method for measuring a glycoprotein according to claim 18, wherein the lectin for normalization is DSA.

20. The method for measuring a glycoprotein according to claim 17 wherein the contacting is performed by contacting the sample, the lectin and a labeled anti- AGP antibody, and forming a complex of the lectin, the AGP, and the labeled anti- AGP antibody,

wherein the measuring is performed by measuring the amount of the label of the complex;

and

quantitatively determining the AGP binding to the lectin.

21. The method for measuring a glycoprotein according to claim 20, wherein the complex is formed using magnetic particles to which the lectin is immobilized.

22. The method for measuring a glycoprotein according to claim 20, wherein the complex is formed using the biotinylated lectin and magnetic particles to which streptavidin or avidin are immobilized.

23. The method for measuring a glycoprotein according to claim 20, wherein the label of the labeled anti- AGP antibody is deglycosylated alkaline phosphatase.

24. The method for measuring a glycoprotein according to claim 20, wherein the anti- AGP antibody is a deglycosylated anti- AGP antibody.

25. The method for measuring a glycoprotein according to claim 20, wherein the measuring is performed by measuring AGP binding to the lectin with the use of a lectin array to which at least the lectin is immobilized and an anti- AGP antibody.

27. A method for examining liver disease, whereby the stage of fiber formation in the liver is determined based on the measured value of AGP obtained by the method for measuring AGP according to claim 17.

28. A method for examining liver disease, whereby whether or not the disease is cirrhosis based on the measured value of AGP obtained by the method for measuring AGP according to claim 17.

29. A method for examining liver disease, whereby a risk of recurrence of hepatic cell carcinoma is determined based on the measured value of AGP obtained by the method for measuring AGP according to claim 17.

30. A method for determining the therapeutic effect of an anti-viral agent in treatment for liver disease, based on the measured value of AGP obtained by the method for measuring AGP according to claim 17.