Method of measuring interaction between biomaterial and sugar chain, method of evaluating biomaterial in sugar chain selectivity, method of screening biomaterial, method of patterning biomaterials, and kits for performing these methods
This invention provides a method of screening or patterning a biomaterial in terms of their specificities to sugar chains by performing real-time and comprehensive measurement of an interaction between sugar chains and the biomaterial concurrently with a very small amount of the biomaterial without labeling. It is a method of measuring an interaction between a biomaterial and a sugar chain(s), the method including: bringing a solution containing the biomaterial in contact with a ligand carrier, the ligand carrier including a support whose surface includes a metal, and a ligand conjugate(s) immobilized independently on the surface, the ligand carrier carrying the ligand conjugate(s) in such a manner that 1 to 500 kinds of the ligand conjugate(s) is immobilized per cm 2 , and each ligand conjugate having a structure in which a sugar chain is bonded with a linker compound having a sulfur atom, and the biomaterial being at least one selected from the groups of proteins, viruses, cells, microorganisms, liposome, and micelles.
1. A method of classifying influenza virus having H3N2 protein by strains, the method comprising:
contacting a solution comprising the influenza virus having H3N2 protein with a ligand carrier,
wherein the ligand carrier comprises a support and a plurality of ligand conjugates,
wherein the support comprises a metal on a surface of the support, and each ligand conjugate of the plurality of ligand conjugates comprises a sugar chain bonded to a linker compound comprising a sulfur atom,
wherein the plurality of ligand conjugates are bonded to the surface via metal-sulfur bonding due to the sulfur atom in the linker compound, and
wherein there are 2 to 500 different kinds of the ligand conjugates, each kind of the ligand conjugate having a different sugar chain, immobilized independently on the ligand carrier per cm 2 , such that the different kinds of the ligand conjugates are bonded to the surface in non-overlapping positions, and wherein the different kinds of ligand conjugates have differing binding specificity to different H3N2 virus strains due to the different sugar chains in the different kinds of ligand conjugates;
measuring interactions between the influenza virus having H3N2 protein and two or more sugar chains of the plurality of ligand conjugates; and
classifying the influenza virus having H3N2 protein by strains, based on differential binding of the influenza virus having H3N2 protein to the different kinds of ligand conjugates,
wherein the classifying further comprises determining where the influenza virus having H3N2 protein interacts with the ligand carrier relative to the positions of the different kinds of ligand conjugates of the plurality of ligand conjugates.
2. The method as set forth in claim 1 , wherein the structure of the linker compound is represented by:
where X is a structure that includes one hydrocarbon chain which has a terminal aromatic amino group with a carbon-nitrogen bond in its main chain, and Y is a hydrocarbon structure having the sulfur atom, and n is not less than 0 but not more than 6.
3. The method as set forth in claim 1 , wherein
the sugar chains are at least two selected from the group consisting of: Galβ1-4Glc, Galβ1-4GlcNAcβ1-6Glc, Neu5Acα2-3Galβ1-4Glc, Neu5Acα2-6Galβ1-4Glc, Neu5Acα2-3Galβ1-3GlcNAcβ1-6Glc, Neu5Acα2-6Galβ1-3GlcNAcβ1-6Glc, Neu5Acα2-3Galβ1-4GlcNAcβ1-6Glc, Neu5Acα2-6Galβ1-4GlcNAcβ1-6Glc, GlcAβ1-3GalNAc4Sβ1-6Glc, GlcAβ1-3GalNAc6Sβ1-6Glc, GlcA2Sβ1-3GalNAc6Sβ1-6Glc, GlcAβ1-3GalNAc4S6Sβ1-6Glc, GlcNS6Sα1-4IdoA2Sα1-6Glc, GlcNSα1-4IdoA2Sα1-6Glc, GlcNS6Sα1-4GlcA2Sβ1-6Glc, and GlcNSα1-4GlcAβ1-6Glc.
4. A kit for performing a method of classifying influenza virus having H 3 N2 protein by strains, the kit comprising:
a solution comprising the influenza virus having H3N2 protein; and
a ligand carrier, wherein the ligand carrier comprises a support and a plurality of ligand conjugates, wherein the support comprises a metal on a surface of the support,
and each ligand conjugate of the plurality of ligand conjugates comprises a sugar chain bonded to a linker compound comprising a sulfur atom,
wherein the plurality of ligand conjugates are bonded to the surface via metal-sulfur bonding due to the sulfur atom in the linker compound and wherein there are 2 to 500 different kinds of the ligand conjugates, each kind of the ligand conjugate having a different sugar chain, immobilized independently on the ligand carrier per cm 2 , such that the different kinds of ligand conjugates are bonded to the surface in non-overlapping positions, and
wherein the different kinds of ligand conjugates have differing binding specificity to the different H3N2 virus strains due to the different sugar chains in the different kinds of ligand conjugates, and
wherein the kit is configured to classify influenza virus having H3N2 protein by strain based on differential binding of the influenza virus to the different kinds of ligand conjugates, wherein the classification comprises determining where the influenza virus interacts with the ligand carrier relative to the positions of the different kinds of ligand conjugates of the plurality of ligand conjugates.
5. The kit as set forth in claim 4 , wherein the sugar chains are at least two selected from the group consisting of: Galβ1-4Glc, Galβ1-4GlcNAcβ1-6Glc, Neu5Acα2-3Galβ1-4Glc, Neu5Acα2-6Galβ1-4Glc, Neu5Acα2-3Galβ1-3GlcNAcβ1-6Glc, Neu5Acα2-6Galβ1-3GlcNAcβ1-6Glc, Neu5Acα2-3Galβ1-4GlcNAcβ1-6Glc, Neu5Acα2-6Galβ1-4GlcNAcβ1-6Glc, GlcAβ1-3GalNAc4Sβ1-6Glc, GlcAβ1-3GalNAc6Sβ1-6Glc, GlcA2Sβ1-3GalNAc6Sβ1-6Glc, GlcAβ1-3GalNAc4S6Sβ1-6Glc, GlcNS6Sα1-4IdoA2Sα1-6Glc, GlcNSα1-4IdoA2Sα1-6Glc, GlcNS6Sα1-4GlcA2Sβ1-6Glc, and GlcNSα1-4GlcAβ1-6Glc.
6. The method of claim 1 , wherein in the measuring the interactions between the influenza virus having H3N2 protein and the two or more sugar chains of the plurality of ligand conjugates, a strength of the interactions between the influenza virus having H3N2 protein and the two or more sugar chains of the plurality of ligand conjugates is measured.
7. The method of claim 1 , wherein in the measuring the interactions between the influenza virus having H3N2 protein and the two or more sugar chains of the plurality of ligand conjugates, the interactions between the influenza virus having H3N2 protein and the two or more sugar chains of the plurality of ligand conjugates are measured concurrently.