Use of heat-resistant biotin-binding protein, and solid support having the protein attached thereto

Abstract

The present invention relates to a solid support having a heat-resistant biotin-binding protein attached thereto. The present invention also relates to the use of the solid support of the present invention having a heat-resistant biotin-binding protein attached thereto. The present invention further relates to technical fields such as purification, concentration, detection and/or capture of a biotin-linked substance by means of a heat-resistant biotin-binding protein. Such a biotin-binding protein used in the solid support of the present invention is heat-resistant and is therefore useful for use in assay systems involving exposure to a temperature of 70° C. or more.

1 . A solid support comprising a heat-resistant biotin-binding protein attached thereto, wherein the heat-resistant biotin-binding protein is selected from the group consisting of:
 a) a protein which comprises the amino acid sequence shown in SEQ ID NO: 2 or SEQ ID NO: 4;   b) a protein which comprises an amino acid sequence comprising deletion, substitution, insertion or addition of one or more amino acids in the amino acid sequence shown in SEQ ID NO: 2 or SEQ ID NO: 4;   c) a protein which comprises an amino acid sequence sharing a sequence identity of at least 60% or more with the amino acid sequence shown in SEQ ID NO: 2 or SEQ ID NO: 4; and   d) a protein which comprises an amino acid sequence encoded by a nucleic acid hybridizable under stringent conditions with the complementary strand of the nucleotide sequence shown in SEQ ID NO: 1 or 3.   
     
     
         2 . The solid support according to  claim 1 , which retains biotin-binding capacity even after heat treatment at 70° C. to 90° C. 
     
     
         3 . The solid support according to  claim 1 , which has biotin-binding capacity even under heating conditions of 70° C. to 90° C. 
     
     
         4 . The solid support according to any one of  claims 1  to  3 , wherein the solid support is composed primarily of a material selected from the group consisting of cellulose, Teflon™, nitrocellulose, agarose, dextran, chitosan, polystyrene, polyacrylamide, polyester, polycarbonate, polyamide, polypropylene, nylon, polydivinylidene difluoride, latex, silica, glass, glass fiber, gold, platinum, silver, copper, iron, stainless steel, ferrite, silicon wafer, polyethylene, polyethyleneimine, polylactic acid, resins, polysaccharides, proteins (e.g., albumin), carbon and combinations thereof. 
     
     
         5 . The solid support according to any one of  claims 1  to  3 , wherein the solid support is selected from the group consisting of beads, magnetic beads, thin films, microtubes, filters, plates, microplates, carbon nanotubes and sensor chips. 
     
     
         6 . Use of the solid support according to  claim 1 , which comprises exposure to heating conditions of 70° C. to 90° C. 
     
     
         7 . A method for separation, concentration, capture, purification and/or detection of a biotin-linked substance, which comprises the following steps:
 1) contacting the solid support according to  claim 1  with a biotin-linked substance, whereby the biotin-linked substance is bound to the solid support;   2) washing off contaminants which are not bound to the solid support; and   3) collecting the biotin-linked substance which is bound to the solid support to thereby separate, concentrate, capture or purify the substance and/or detect the substance   wherein at least one of the above steps is accomplished under heating conditions of 70° C. to 90° C.   
     
     
         8 . The method according to  claim 7 , wherein the heating conditions are set to 75° C. to 85° C. 
     
     
         9 . The method according to  claim 7 , wherein the biotin-linked substance is a biotin-linked nucleic acid.