(1-3)-beta-D-glucan binding domain protein, measuring method using the substance and assay kit

Abstract

A (1→3)-β-D-glucan binding protein, a fluorescence-labeled (1→3)-β-D-glucan binding domain protein, a (1→3)-β-D-glucan measuring agent comprising the same, a method for measuring (1→3)-β-D-glucan using the same, and a (1→3)-β-D-glucan assay kit comprising the same.

What is claimed is:  
     
         1 . A fluorescence-labeled (1→3)-β-D-glucan binding domain protein, which comprises: 
 a protein comprising the amino acid sequence represented by SEQ ID NO:2; or  
 a protein comprising an amino acid sequence represented by SEQ ID NO:2 in which a substitution, deletion, insertion, addition or transposition of at least one amino acid residue is made, having a molecular weight of from 10 kDa to 40 kDa, and being capable of binding to (1→3)-β-D-glucan,  
 wherein a fluorescent material is bound to the protein.  
 
     
     
         2 . A (1→3)-β-D-glucan measuring agent, which comprises the fluorescence-labeled (1→3)-β-D-glucan binding domain protein according to  claim 1 .  
     
     
         3 . The agent according to  claim 2 , which further comprises a divalent cation.  
     
     
         4 . The agent according to  claim 3 , wherein the divalent cation is an alkaline earth metal ion.  
     
     
         5 . A method for measuring (1→3)-β-D-glucan, which comprises: 
 binding the fluorescence-labeled (1→3)-β-D-glucan binding domain protein according to  claim 1  to (1→3)-β-D-glucan in a sample;  
 detecting a change in a degree of fluorescence polarization caused by the binding; and  
 correlating a changed amount of the degree of fluorescence polarization with a concentration of (1→3)-β-D-glucan in the sample.  
 
     
     
         6 . The method according to  claim 5 , wherein the fluorescence-labeled (1→3)-β-D-glucan binding domain protein is bound to the (1→3)-β-D-glucan in the presence of a divalent cation.  
     
     
         7 . A (1→3)-β-D-glucan assay kit, which comprises the fluorescence-labeled (1→3)-β-D-glucan binding domain protein according to  claim 1 .  
     
     
         8 . The kit according to  claim 7 , which further comprises a divalent cation.  
     
     
         9 . The kit according to  claim 8 , wherein the divalent cation is an alkaline earth metal ion.  
     
     
         10 . A protein which consists of: 
 a protein consisting of the amino acid sequence represented by SEQ ID NO:2; or    a protein consisting of an amino acid sequence represented by SEQ ID NO:2 in which a substitution, deletion, addition or transposition of at least one amino acid residue is made, having a molecular weight of from 10 kDa to 40 kDa, and being capable of binding to (1→3)-β-D-glucan.    
     
     
         11 . A DNA encoding a protein which consists of: 
 a protein consisting of the amino acid sequence represented by SEQ ID NO:2; or    a protein consisting of an amino acid sequence represented by SEQ ID NO:2 in which a substitution, deletion, addition or transposition of at least one amino acid residue is made, having a molecular weight of from 10 kDa to 40 kDa, and being capable of binding to (1→3)-β-D-glucan.    
     
     
         12 . A DNA which consists of: 
 a DNA consisting of the nucleotide sequence represented by SEQ ID NO:1; or    a DNA which hybridizes to a nucleotide sequence complementary to the nucleotide sequence represented by SEQ ID NO:1 under stringent conditions, and which encodes a protein having a molecular weight of from 10 kDa to 40 kDa and being capable of binding to (1→3)-β-D-glucan.    
     
     
         13 . A method for detecting mycosis, which comprises: 
 binding the fluorescence-labeled (1→3)-β-D-glucan binding domain protein according to  claim 1  to (1→3)-β-D-glucan in a sample;    detecting a change in a degree of fluorescence polarization caused by the binding; and    correlating a changed amount of the degree of fluorescence polarization with existence of mycosis.