# US 11,078,254 B2 · Method for immobilizing lectin > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,078,254 B2 - Document ID: US11078254B2 - Status: Granted - Kind: B2 - Application number: 16/604,565 - Filed: 2018-04-11 - Published: 2020-04-23 - Granted: 2021-08-03 - Earliest claimed: 2017-04-11 - Assignees: National Institute of Advanced Industrial Science and Technology; Nissan Chemical Corporation; National University Corporation Kyoto Institute of Technology - Applicants: National Institute of Advanced Industrial Science and Technology; Nissan Chemical Corporation; National University Corporation Kyoto Institute of Technology - Inventors: Hiroaki Tateno; Junko Katayama; Kazutaka Matoba; Yoichi Kumada - Web page: https://patentgrep.com/patent/US11078254B2 ## CPC - C07K 14/4726: {Lectins} - C07K 14/7056: {Lectin superfamily, e.g. CD23, CD72} - C07K 7/06: having 5 to 11 amino acids - C07K 7/08: having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - C07K 14/195: from bacteria - C07K 17/00: Carrier-bound or immobilised peptides (carrier-bound or immobilised enzymes C12N11/00); Preparation thereof - C07K 17/08: the carrier being a synthetic polymer - C07K 17/14: Peptides being immobilised on, or in, an inorganic carrier - C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} - C12N 15/09: Recombinant DNA-technology - C12N 15/62: DNA sequences coding for fusion proteins - G01N 33/543: with an insoluble carrier for immobilising immunochemicals - G01N 33/54353: {with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393)} - G01N 33/545: Synthetic resin - G01N 33/551: the carrier being inorganic - G01N 33/566: using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} - G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells} - C07K 2319/00: Fusion polypeptide - C07K 2319/20: containing a tag with affinity for a non-protein ligand - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation - G01N 2333/42: Lectins, e.g. concanavalin, phytohaemagglutinin - G01N 2333/4724: Lectins - G01N 2400/02: involving antibodies to sugar part of glycoproteins - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Provided is a highly sensitive and less expensive lectin-immobilized base material (for example, a lectin plate), such as lectin-immobilized base material having stable qualities and being able to be sufficiently washed after a target sugar chain-containing antigen binds thereto. Further provided is a method for immobilizing lectin to a base material therefor. Particularly provided are: a method whereby a lectin-peptide fusion, in which a peptide capable of adsorbing to a base material surface such as a polystyrene (PS) tag is fused with the N-terminal side or C-terminal side of lectin capable of recognizing a target sugar chain, is immobilized on the peptide side to a base material; and a lectin-immobilized base material produced by this method. By using the lectin-immobilized base material, a target sugar chain-containing antigen can be highly sensitively and evenly measured and, moreover, target sugar chain-containing cells, etc. can be separated (concentrated and harvested). ## Claims ### Claim 1 A lectin-peptide fusion, comprising: a peptide capable of adsorbing to a base material surface; and a lectin capable of recognizing a target sugar chain, wherein the peptide is provided on an N-terminal side or a C-terminal side of the lectin, wherein the peptide is a PS peptide including a PSI tag having an amino acid sequence of SEQ ID NO: 2, or a PSS tag having an amino acid sequence of SEQ ID NO: 9. ### Claim 2 (depends on 1) The fusion according to claim 1, wherein the lectin is a lectin selected from the group consisting of a BC2LCN lectin, a rACG lectin, a rPSL1a lectin, a rLSLN lectin, a rDiscoidin I lectin, a rDiscoidin II lectin, a rCGL2 lectin, a rSRL lectin, a rF17AG lectin, a rGRFT lectin, a rOrysata lectin, a rCalsepa lectin, a rBC2LA lectin, a rAAL lectin, a rPAIIL lectin, a rRSIIL lectin, a rPPL lectin, a rCNL lectin, a rPAIL lectin, a rABA lectin, a rMOA lectin, a rPALa lectin, a rGal3CS lectin, a rMpL lectin, a rAAL2 lectin, a rBambL lectin, and a rPVL lectin. ### Claim 3 (depends on 1) The fusion according to claim 1, wherein the lectin is a BC2LCN lectin having the amino acid sequence of SEQ ID NO: 24. ### Claim 4 A polynucleotide encoding a lectin-peptide fusion, wherein the lectin-peptide fusion comprises a peptide capable of adsorbing to a base material surface; and a lectin capable of recognizing a target sugar chain, wherein the peptide is provided on an N-terminal side or a C-terminal side of the lectin, wherein the peptide is a PS peptide including a PSI tag having an amino acid sequence of SEQ ID NO:2, or a PSS tag having an amino acid sequence of SEQ ID NO:9. ### Claim 5 (depends on 4) A vector capable of expressing a lectin-peptide fusion, comprising the polynucleotide of claim 4. ### Claim 6 (depends on 1) An immobilized lectin-peptide fusion, comprising: the lectin-peptide fusion according to claim 1; and a base material, wherein the peptide side in the lectin-peptide fusion is immobilized on the base material. ### Claim 7 (depends on 1) A base material, on which the lectin-peptide fusion according to claim 1 is immobilized. ### Claim 8 (depends on 7) A method for measuring or isolating a target sugar chain-containing antigen, comprising a process of bringing a sample containing a target sugar chain-containing antigen into contact with the base material according to claim 7. ### Claim 9 (depends on 8) The method according to claim 8, further comprising a process of overlaying an antibody capable of recognizing the target sugar chain-containing antigen. ### Claim 10 (depends on 9) The method according to claim 9, wherein in the process of overlaying an antibody capable of recognizing the target sugar chain-containing antigen, a blocking agent is contained in a diluent for a base material on which a lectin-peptide fusion is immobilized; and/or in a diluent for an antibody. ### Claim 11 (depends on 9) The method according to claim 9, wherein the target sugar chain-containing antigen is a sugar chain-containing antigen contained in a solution containing contaminants derived from a test sample. ### Claim 12 (depends on 7) A method for concentrating or isolating a cell having a target sugar chain or a glycoconjugate, comprising: a process of allowing a sample containing a cell having a target sugar chain on a surface of the cell or a glycoconjugate having a target sugar chain to adsorb to the base material according to claim 7; and a process of harvesting a target sugar chain-containing substance. ### Claim 13 (depends on 7) A kit or device for measuring or isolating a target sugar chain-containing antigen, comprising: the base material according to claim 7. ### Claim 14 (depends on 13) The kit or device according to claim 13, further comprising an antibody capable of recognizing the target sugar chain-containing antigen. ### Claim 15 (depends on 1) A method for producing a base material on which a lectin-peptide fusion is immobilized, comprising a process of bringing the lectin-peptide fusion according to claim 1 into contact with a base material. ### Claim 16 (depends on 15) The production method according to claim 15, wherein the process of bringing the lectin-peptide fusion into contact with a base material is performed in a buffer solution. ### Claim 17 (depends on 16) The production method according to claim 16, wherein the buffer solution has a pH of 6.5 to 7.5. ### Claim 18 (depends on 16) The production method according to claim 16, wherein the buffer solution has a salt concentration of 0.10 to 0.20 M.