Method of binding proteins to carriers by making use of tamavidins
The present invention provides a method of binding a protein to a carrier in such a way that the protein is not impaired in its function but can be allowed to act more efficiently than when it is bound directly. The method of the present invention for binding a protein to a carrier comprises: preparing a biotin-bound carrier; preparing a fusion protein having the protein bound to a tamavidin; and binding the protein to the carrier via tamavidin-biotin bonds.
1. A method of binding a protein to a carrier, which comprises:
providing a biotin-bound carrier;
preparing a fusion protein having the protein bound to a tamavidin; and
binding the protein to the carrier via tamavidin-biotin bonds,
wherein the tamavidin is selected from:
(a) a protein consisting of an amino acid sequence sharing an identity of 90% or more with SEQ ID NO: 2 or SEQ ID NO: 4 and having biotin-binding activity; or
(b) a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4; or
(c) a protein consisting of an amino acid sequence encoded by a nucleic acid hybridizable under stringent conditions (0.5% SDS, 65° C., 0.2×SSC) with a strand complementary to the base sequence of SEQ ID NO: 1 or SEQ ID NO: 3 and having biotin-binding activity.
2. The method according to claim 1 , wherein the protein is selected from the group consisting of antibodies or fragments thereof, antigenic proteins, enzymes, lectins, peptides, protein A, protein G, and protein L.
3. The method according to claim 1 , wherein the carrier is selected from the group consisting of beads, magnetic beads, thin films, microtubes, filters, plates, microplates, carbon nanotubes, and sensor chips.
4. The method according to claim 1 , wherein the tamavidin and the protein are bound via a linker to constitute the fusion protein.
5. The method according to claim 1 , wherein the tamavidin and the protein are bound via a linker consisting of six or more amino acids to constitute the fusion protein.
6. The method according to claim 1 , wherein the fusion protein further has a leader sequence bound thereto.
7. The method according to claim 1 , wherein biotin and the carrier are bound via a linker greater than 13.5 Å in length.
8. A tamavidin fused protein-bound carrier in which a fusion protein having a protein bound to a tamavidin is bound to a biotin-bound carrier via tamavidin-biotin bonds,
wherein the tamavidin is selected from:
(a) a protein consisting of an amino acid sequence sharing an identity of 90% or more with SEQ ID NO: 2 or SEQ ID NO: 4 and having biotin-binding activity; or
(b) a protein consisting of the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4; or
(c) a protein consisting of an amino acid sequence encoded by a nucleic acid hybridizable under stringent conditions (0.5% SDS, 65° C., 0.2×SSC) with a strand complementary to the base sequence of SEQ ID NO: 1 or SEQ ID NO: 3 and having biotin-binding activity,
wherein the tamavidin fused protein-bound carrier is prepared by the method of claim 1 for binding a protein to a carrier.