# US 8,785,134 B2 · Glycoprotein vesicles and their methods of use > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,134 B2 - Document ID: US8785134B2 - Status: Granted - Kind: B2 - Application number: 12/268,333 - Filed: 2008-11-10 - Published: 2012-06-28 - Granted: 2014-07-22 - Earliest claimed: 2008-11-10 - Assignees: The MITRE Corporation - Applicants: Elaine H. Mullen - Inventors: Elaine H. Mullen - Web page: https://patentgrep.com/patent/US8785134B2 ## CPC - G01N 21/82: producing a precipitate or turbidity - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01N 21/94: Investigating contamination, e.g. dust (G01N21/85 takes precedence) - G01N 2021/7786: {Fluorescence} - G01N 2021/825: {Agglutination} - 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 are methods of collecting, detecting and altering cells and molecular entities using glycoprotein micelles and vesicles. Glycoprotein vesicles comprising a glycoprotein micelle, at least a monolayer of lectin and/or a monolayer of biologically active glycoproteins are also provided. The invention further provides methods of detecting protein glycosylation using the vesicles of the invention. ## Claims ### Claim 1 A glycoprotein vesicle-containing suspension in an aqueous polar medium, wherein the glycoprotein vesicle comprises: (a) a glycoprotein micelle comprising a hydrophobic interior and a polar exterior, wherein the glycoprotein's carbohydrate moieties are on the polar exterior and are oriented toward the aqueous polar medium in which the vesicle is suspended; and (b) at least one monolayer of a lectin non-covalently or covalently bound to the carbohydrate moieties that are on the polar exterior of the micelle of part (a), wherein the at least one monolayer of a lectin of part (b) is on the outer surface of the vesicle and is exposed to the aqueous polar medium that contains the vesicle, wherein the vesicles are not biological cells and are not agglutinated, and wherein the suspension does not contain non-bound lectin or non-bound glycoprotein. ### Claim 2 (depends on 1) A method of collecting one or more glycoprotein contaminants or glycan-coated pathogens from an aqueous environment comprising (a) contacting the glycoprotein vesicle-containing suspension of claim 1 with an aqueous solution or suspension containing the one or more glycoprotein contaminants or glycan-coated pathogens, (b) allowing the vesicles to agglutinate with the glycoprotein contaminants or glycan-coated pathogens, and (c) collecting the agglutinated vesicles to collect the glycoprotein contaminants or glycan-coated pathogen from the aqueous solution or suspension, wherein the presence of agglutinated vesicles indicates that one or more of the glycoprotein contaminants or glycan-coated pathogens have been collected from the aqueous environment. ### Claim 3 (depends on 2) The method of claim 2, wherein the one or more glycoprotein contaminants or glycan-coated pathogens is the one or more glycoprotein contaminants. ### Claim 4 (depends on 2) The method of claim 2, wherein the one or more glycoprotein contaminants or glycan-coated pathogens is the one or more glycan-coated pathogens. ### Claim 5 (depends on 1) The suspension of claim 1, wherein the lectin is selected from the group consisting of a plant lectin, an animal lectin, a bacterial lectin and a viral lectin. ### Claim 6 (depends on 1) The suspension of claim 1, wherein the lectin is a bacterial or viral lectin. ### Claim 7 A method of detecting glycosylation of a protein comprising (a) forming transparent micelles of the protein, (b) contacting the transparent micelles of (a) with at least one labeled lectin; and (c) detecting a signal generated from the circumference of the transparent micelles and from the center of the transparent micelles, wherein a stronger signal at the circumference of the transparent micelles than in the center of the transparent micelles indicates that the protein is glycosylated. ### Claim 8 (depends on 7) The method of claim 7, wherein the lectin is labeled with a fluorophore. ### Claim 9 (depends on 8) The method of claim 8, wherein the fluorophore is fluorescein. ### Claim 10 (depends on 7) The method of claim 7, wherein the lectin binds to a conserved core of an N-linked oligosaccharide. ### Claim 11 (depends on 10) The method of claim 10, wherein the lectin is concanavalin A. ### Claim 12 A glycoprotein vesicle-containing suspension in an aqueous polar medium, wherein the glycoprotein vesicle comprises: (a) a glycoprotein micelle comprising a hydrophobic interior and a polar exterior, wherein the glycoprotein's carbohydrate moieties are on the polar exterior and are oriented toward the aqueous polar medium in which the vesicle is suspended; (b) at least one monolayer of a lectin non-covalently or covalently bound to the carbohydrate moieties that are on the polar exterior of the micelle of part (a); and (c) at least one monolayer of a glycoprotein covalently or non-covalently bound to the lectin layer such that the at least one monolayer of lectin of part (b) is between the carbohydrate moieties that are on the exterior of the micelle of part (a) and the at least one monolayer of a glycoprotein of part (c), wherein the at least one monolayer of a glycoprotein of part (c) is on the outer surface of the vesicle and is exposed to the aqueous polar medium that contains the vesicle; wherein the vesicles are not biological cells and are not agglutinated, and wherein the suspension does not contain non-bound lectin or non-bound glycoprotein. ### Claim 13 (depends on 12) A method of modifying the carbohydrate portion of a glycoprotein comprising contacting the vesicle of claim 12 with a glycosyl transferase in the presence of at least one carbohydrate. ### Claim 14 (depends on 12) The suspension of claim 12, wherein the glycoprotein in the glycoprotein micelle comprises a glycoprotein selected from the group consisting of ovomucoid, yeast invertase, ovalbumin, ovotransferrin, transferrin, and uromodulin. ### Claim 15 (depends on 12) The suspension of claim 12, wherein the glycoprotein in the at least one monolayer of a glycoprotein is an enzyme inhibitor, an enzyme, a co-enzyme, mucin, uromodulin, a hormone, an apo lipoprotein, an antibody, or a metal binding glycoprotein. ### Claim 16 (depends on 12) The suspension of claim 12, wherein the lectin is selected from the group consisting of a plant lectin, an animal lectin, a bacterial lectin and a viral lectin. ### Claim 17 (depends on 1, 12) A method of detecting a biological contaminant in an aqueous solution comprising: (a) contacting the suspension of claim 1 or claim 12 with the aqueous solution; and (b) detecting whether the vesicles agglutinate after the contacting of the vesicles with the aqueous solution, wherein agglutination indicates the presence of the contaminant. ### Claim 18 (depends on 17) The method of claim 17, wherein the contaminant is a pathogen or a biotoxin. ### Claim 19 (depends on 17) The method of claim 17, wherein the biological contaminant is glycosylated.