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US 8,945,509 B2Granted
Magnetic beads having surface glycoconjugates and use thereof
AssigneeLife Technologies Corporation
InventorsAldrich Lau, Robert Eason, Maxim Brevnov, Handong Li, Kevin Hacker
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Abstract
Magnetic beads that include polyvalent ligands comprising various carbohydrates are described. Methods for fabricating such magnetic beads are also provided as well as methods of their use to capture and enrich pathogen cell population for subsequent culture, lysis and identification.
Claims33
A composition, comprising:
- (a) a paramagnetic bead;
- (b) a plurality of hydrophilic copolymer bridges, each bridge being covalently bonded to the paramagnetic bead;
- and
- (c) a plurality of carbohydrates, each carbohydrate being covalently bonded to the same or different hydrophilic copolymer bridge by forming a glycoconjugate with the respective hydrophilic copolymer, wherein each hydrophilic polymer bridge is bonded to the paramagnetic bead via a link formed by an amino group, wherein at least one hydrophilic copolymer is different from at least one other hydrophilic copolymer.
1 more element
A method for fabricating a paramagnetic bead composition, the method comprising:
- (a) immobilizing a plurality of functionalized hydrophilic copolymers on the surface of a paramagnetic bead;
- and
- (b) bonding at least one carbohydrate to each of at least two hydrophilic copolymers, wherein each of the functionalized hydrophilic copolymers includes a plurality of functional groups, comprising a first functional group and at least one second functional group, to thereby obtain the paramagnetic bead composition, wherein the step of bonding comprises reacting the plurality of functionalized hydrophilic copolymers with the carbohydrate(s), wherein at least two glycoconjugates are formed between at least one carbohydrate and the hydrophilic copolymers, wherein the step of immobilizing comprises reacting the plurality of functionalized hydrophilic copolymers with the paramagnetic bead, wherein a covalent linkage is formed between the paramagnetic bead and each hydrophilic copolymer.
A composition, comprising:
- (a) a paramagnetic bead;
- (b) a plurality of hydrophilic copolymer bridges, each bridge being covalently bonded to the paramagnetic bead;
- and
- (c) a plurality of carbohydrates, each carbohydrate being covalently bonded to the same or different hydrophilic copolymer bridge by forming a glycoconjugate with the respective hydrophilic copolymer, wherein each hydrophilic polymer bridge is bonded to the paramagnetic bead via a link formed by an amino group,
- wherein the hydrophilic copolymer bridges are formed of functionalized hydrophilic copolymers selected from the group consisting of functionalized acrylates, poly(alkylene glycols), alkoxy poly(alkylene glycols), copolymers of methylvinyl ether and maleic acid, urethanes, ethyleneimines, polyurethane-polyether copolymers, copolymers having units derived from vinyl alcohol, N-vinyl lactams, vinyl pyrrolidone, amides, maleic anhydride, styrenesulfonates, vinylsulfonic acid, vinylsulfonates, N-vinylamides or 3-hydroxybutyric acid, and,
- wherein the functionalized acrylates are selected from the group consisting of functionalized copolymers having units derived from acrylic acid, methacrylic acid, 2-hydroxyalkyl acrylate, 2-hydroxyalkylmethacrylate, acrylamides, methacrylamides, epoxy-acrylates, and,
- wherein each of the functionalized hydrophilic copolymers includes a plurality of functional groups, comprising:
- (a) a first functional group to form a covalent linkage between the magnetic bead and the respective hydrophilic copolymer bridge;
- and
- (b) at least one second functional group, to attach at least one carbohydrate to the hydrophilic copolymer by forming glycoconjugate(s) between the carbohydrate(s) and the hydrophilic copolymer, and
- wherein in the plurality of functional groups at least one functional group is different from at least one other functional group.
8 more elements
A composition, comprising:
- (a) a paramagnetic bead;
- (b) a plurality of hydrophilic copolymer bridges, each bridge being covalently bonded to the paramagnetic bead;
- and
- (c) a plurality of carbohydrates, each carbohydrate being covalently bonded to the same or different hydrophilic copolymer bridge by forming a glycoconjugate with the respective hydrophilic copolymer, wherein each hydrophilic polymer bridge is bonded to the paramagnetic bead via a link formed by an amino group,
- wherein at least one carbohydrate is different from at least one other carbohydrate.
2 more elements
A composition, comprising:
- (a) a paramagnetic bead;
- (b) a plurality of hydrophilic copolymer bridges, each bridge being covalently bonded to the paramagnetic bead;
- and
- (c) a plurality of carbohydrates, each carbohydrate being covalently bonded to the same or different hydrophilic copolymer bridge by forming a glycoconjugate with the respective hydrophilic copolymer, wherein each hydrophilic polymer bridge is bonded to the paramagnetic bead via a link formed by an amino group,
- wherein each hydrophilic copolymer has the Hildebrand solubility parameter δ of at least about 10 MPa1/2.
2 more elements
A method for fabricating a paramagnetic bead composition, the method comprising:
- (a) immobilizing a plurality of functionalized hydrophilic copolymers on the surface of a paramagnetic bead;
- and
- (b) bonding at least one carbohydrate to each of at least two hydrophilic copolymers, wherein each of the functionalized hydrophilic copolymers includes a plurality of functional groups, comprising a first functional group and at least one second functional group, to thereby obtain the paramagnetic bead composition, wherein the step of bonding comprises reacting the plurality of functionalized hydrophilic copolymers with the carbohydrate(s), wherein at least two glycoconjugates are formed between at least one carbohydrate and the hydrophilic copolymers, wherein in the plurality of functional groups at least one functional group is different from at least one other functional group.
A method for fabricating a paramagnetic bead composition, the method comprising:
- (a) immobilizing a plurality of functionalized hydrophilic copolymers on the surface of a paramagnetic bead;
- and
- (b) bonding at least one carbohydrate to each of at least two hydrophilic copolymers, wherein each of the functionalized hydrophilic copolymers includes a plurality of functional groups, comprising a first functional group and at least one second functional group, to thereby obtain the paramagnetic bead composition, wherein the step of bonding comprises reacting the plurality of functionalized hydrophilic copolymers with the carbohydrate(s), wherein at least two glycoconjugates are formed between at least one carbohydrate and the hydrophilic copolymers, wherein at least two carbohydrates are bonded and at least one individual carbohydrate is different from at least one other individual carbohydrate.
Dependencies are inferred from the claim text.