Method for measuring β-glucan, and β-glucan-binding protein for use in the method

A kit for measuring β-glucan comprising the followings as constituents:

• (1) a reagent containing a protein 1 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any of the SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8 and SEQ ID NO:
• 10, and having a β-glucan binding activity (hereinafter, abbreviated as “βG-binding protein 1”);
• and
• (2) a reagent containing a protein 2 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of the SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity. (hereinafter, abbreviated as “βG-binding protein 2”),
• wherein the βG-binding protein 2 is labeled with a labeling substance.

An isolated protein comprising an amino acid sequence identical to an amino acid sequence shown in any one of SEQ ID NO:

• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity.

A method for measuring βG comprising:

• (1) contacting a sample with
• βG-binding protein 1 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity (hereinafter, abbreviated as “βG-binding protein 1”), wherein the βG-binding protein 1 is immobilized to an insoluble carrier, and
• βG-binding protein 2 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity (hereinafter, abbreviated as “βG-binding protein 2”), to form a complex of the βG-binding protein 1, βG in the sample and the βG-binding protein 2,
• (2) measuring a quantity of the complex by a turbidimetric method, a nephelometric method, or a latex aggregation method;
• and
• (3) determining a βG concentration in the sample based on the quantity measured in (2).

A method for measuring βG comprising:

• (1) contacting a sample with
• βG-binding protein 1 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity, and
• βG-binding protein 2 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity, to form a complex of the βG-binding protein 1, βG in the sample and the βG-binding protein 2,
• wherein the βG-binding protein 1 and βG-binding protein 2 are different each other, and
• wherein at least one of the βG-binding protein 1 and the βG-binding protein 2 comprises an amino acid sequence shown in SEQ ID NO:
• 4 or SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in SEQ ID NO:
• 4, or SEQ ID NO:
• 10, and having a β-glucan binding activity and peroxidase activity;
• (2) measuring quantity of the complex by measuring a peroxidase activity derived from the βG-binding protein 1 and/or βG-binding protein 2;
• and
• (3) determining a βG concentration in the sample based on the quantity.

A kit for measuring β-glucan comprising the followings as constituents:

• (1) a reagent containing a protein 1 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8m, and SEQ ID NO:
• 10, and having a β-glucan binding activity, wherein the βG-binding protein 1 is immobilized to an insoluble carrier, and
• (2) a reagent containing a protein 2 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity.

A kit for measuring βG comprising the followings as constituents:

• (1) a reagent contacting a protein 1 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity, and
• (2) a reagent containing a protein 2 comprising an amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10,or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in any one of SEQ ID NO:
• 4, SEQ ID NO:
• 6, SEQ ID NO:
• 8, and SEQ ID NO:
• 10, and having a β-glucan binding activity,
• wherein the βG-binding protein 1 and βG-binding protein 2 are different each other, and
• wherein at least one of the βG-binding protein 1 and the βG-binding protein 2 comprises an amino acid sequence shown in SEQ ID NO:
• 4 or SEQ ID NO:
• 10, or an amino acid sequence having 95% or more of homology to the amino acid sequence shown in SEQ ID NO:
• 4, or SEQ ID NO:
• 10, and having a β-glucan binding activity and peroxidase activity.