# US 2020/0399401 A1 · Bioconjugates of Antibodies and Fuctionalized Magnetic Nanoparticles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399401 A1 - Document ID: US20200399401A1 - Status: Published application - Kind: A1 - Application number: 16/977,623 - Filed: 2019-02-25 - Published: 2020-12-24 - Earliest claimed: 2018-03-02 - Assignees: UniversitäT ZüRich; ETH Zurich - Applicants: Universität Zürich; ETH Zurich - Inventors: Antoine Florent Herzog; Wendelin Jan Stark; Martin Zeltner; Beatrice Beck-Schimmer; Anja Zabel - Web page: https://patentgrep.com/patent/US20200399401A1 ## CPC - A61K 47/6935: {the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol} - C07K 17/14: Peptides being immobilised on, or in, an inorganic carrier - A61K 39/39558: {against tumor tissues, cells, antigens} - A61K 47/6929: {the form being a nanoparticle, e.g. an immuno-nanoparticle} - A61K 49/0093: {Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle} - C07K 16/30: from tumour cells - G01N 33/553: Metal or metal coated - G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells} - A61K 2039/505: {comprising antibodies} - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - 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) - 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 The present invention relates to the field of antibodies useful in therapy and diagnosis. It specifically discloses bioconjugates comprising one or more antibodies bound to functionalized, magnetic nanoparticles. These bioconjugates are useful in cancer therapy, particularly for removing circulating tumor cells. The invention further pertains to specific functionalized, magnetic nanoparticles and to the manufacturing of such bioconjugates and such functionalized, magnetic nanoparticles. ## Claims ### Claim 1 A Bioconjugate containing a nanoparticle of the core shell type and one or more antibodies immobilized thereon, wherein: said core contains a metal or alloy having soft magnetic properties and said shell contains one or more graphene layers, wherein the outermost layer is functionalized by one or more of the groups according to formula (I): wherein (Z)m represents a spacer selected from polyglycidol with m repeating units; m is an integer between 10 and 30; FG represents independent from each other a functional group selected from OH, COOH, COOR, and CO(NH)R; R represents C1-C4 alkyl; and n is an integer between 6 and 100. ### Claim 2 (depends on 1) The Bioconjugate according to claim 1, wherein said antibody is a monoclonal antibody and/or specifically binds to circulating tumor cells, preferably wherein said antibody is an anti-EpCam antibody; and/or wherein said core contains, particularly consist of, Co, Fe, Ni or its alloys; and/or wherein in groups according to formula (I) FG represents OH and n is an integer between 10 and 60. ### Claim 3 (depends on 1) The Bioconjugate according to claim 1, wherein said immobilization is effected by covalent bonding, preferably wherein said covalent bonding comprises at least one covalent bond between one functional group FG and one antibody AB. ### Claim 4 (depends on 3) The Bioconjugate according to claim 3, wherein said covalent bonding comprises at least one covalent bond between one functional group FG and a coupling group of formula (II) and at least one covalent bond between said coupling group of formula (II) and one antibody AB; wherein said coupling group is of formula (II) wherein R2 represents a C1-6 alkandiyl, C2-6 alkendiyl, C3-6 cycloalkyl, phenyl X1, represents O, NR1, X2 represents O, NR1, R1 represents C1-C4 alkyl; FG represents the functional group as defined in formula (I) and (AB) represents said antibody. ### Claim 5 (depends on 1) The Bioconjugate according to claim 1, wherein: said bioconjugate contains one nanoparticle and 1-100 antibodies immobilized thereon; and/or said bioconjugate has an average diameter between 30-100 nm; said core has a diameter between 10-200 nm; and/or said shell has a thickness of 0.3-10 nm. ### Claim 6 (depends on 1) The Bioconjugate according to claim 1, for the use as a pharmaceutical, particularly for use in therapy. ### Claim 7 (depends on 1) The Bioconjugate according to claim 1 for the treatment of cancer, particularly for removing circulating tumor cells from blood. ### Claim 8 A Nanoparticle of the core shell type, wherein: said core contains a metal or alloy having soft magnetic properties and said shell contains one or more graphene layers and where the outermost layer is functionalized by one or more of the groups according to formula (I): wherein (Z)m represents a spacer selected from polyglycidol with m repeating units; m is an integer between 10 and 30; FG represents independent from each other a functional group selected from OH, COOH, COOR, and CO(NH)R; R represents C1-C4 alkyl; and n is an integer between 6 and 100. ### Claim 9 (depends on 8) The Nanoparticle according to claim 8, wherein (Z)m is a spacer selected from polyglycidol with m repeating units; and m is an integer between 10 and 30; and FG represents OH and n is an integer between 10 and 60. ### Claim 10 (depends on 1, 8) A method for manufacturing nanoparticles according to claim 8, said method comprising the steps of: a) providing a nanoparticle of the core shell type, wherein: said core contains a metal or alloy having soft magnetic properties and said shell contains one or more graphene layers which are functionalized by one or more of the groups according to formula (III): wherein R3 represents a direct bond or C1-6 alkandiyl, C2-6 alkendiyl, C3-6 cycloalkyl; preferably 1,2-ethandiyl; b) providing a compound of formula (IV) wherein o is an integer between 1 and 4, preferably 1, p is an integer between 1 and 4, preferably 1, FG is as defined in claim 1; c) subjecting compounds of formulae (II) and (IV) to a ring opening polymerization, optionally in the presence of a diluent, optionally in the presence of a reaction aid to thereby obtain said nanoparticles. ### Claim 11 (depends on 10) A nanoparticle, obtained by or obtainable by a method according to claim 10. ### Claim 12 (depends on 1, 8, 9, 11) A method for manufacturing a bioconjugate according to claim 1, said method comprising the steps of: a) providing nanoparticles as defined in any of claims 8 to 9 and 11 in a diluent; b) providing antibodies in a diluent; c) contacting said nanoparticles with said antibodies to thereby obtain said bioconjugate; whereby either said nanoparticles or said antibodies are activated prior to said contacting step. ### Claim 13 (depends on 1, 4, 8, 9, 11) A method for manufacturing a bioconjugate according to claim 1, said method comprising the steps of: a) providing nanoparticles as defined in any of claims 8 to 9 and 11 in a diluent; providing a coupling agent of formula (IIa) or (IIb), optionally in a diluent, whereby X1, X2, R2, are as defined in claim 4 and LG1, LG2 are leaving groups, preferably hydroxyl; b) providing antibodies in a diluent; c) contacting said nanoparticles with said antibodies and said coupling agent (IIa) or (IIb) to thereby obtain said bioconjugate; d) optionally purifying the thus obtained antibody; whereby said nanoparticles are first contacted with said coupling agent and the thus obtained nanoparticle is contacted with said antibody, optionally the presence of reactions aids. ### Claim 14 (depends on 1, 4, 8, 9, 11) A method for manufacturing a bioconjugate according to claim 1, said method comprising the steps of: a) providing nanoparticles as defined in any of claims 8 to 9 and 11 in a diluent; b) providing antibodies in a diluent; providing a coupling agent of formula (IIa) or (IIb), optionally in a diluent, whereby X1, X2, R2, are as defined in claim 4 and LG1, LG2 are leaving groups, preferably hydroxyl; c) contacting said nanoparticles with said antibodies and said coupling agent (IIa) or (IIb) to thereby obtain said bioconjugate; d) optionally purifying the thus obtained antibody; whereby said antibodies are first contacted with said coupling agent and the thus obtained modified antibodies are contacted with said nanoparticles, optionally the presence of reactions aids. ### Claim 15 (depends on 12) A bioconjugate, obtained or obtainable by a method according to claim 12.