Universal functionalized beads and method of making same
AssigneeRAN Biotechnologies, Inc.
InventorsRobert E. Lintner, Roger A. Nassar
Life of this patent
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Abstract
The present invention is generally directed to beads, methods of making said beads, and methods of functionalizing said beads with one or more functional groups to form functionalized beads, such as universal functionalized beads, wherein the functional group can be a chemical group. In some embodiments, the beads can be gel beads.
Claims25
A method of making gel beads, the method comprising:
- suspending a first fluid sample in a second fluid sample to form a plurality of fluidic droplets, wherein the first fluid sample comprises a plurality of first species;
- and
- gelating the fluidic droplets to form a plurality of first gel beads comprising the first species,
- wherein one or more of the first species have one or more chemical functionalities.
A method of making gel beads, the method comprising:
- suspending a first fluid sample in a second fluid sample to form a plurality of fluidic droplets, wherein the first fluid sample comprises a plurality of species and solid templating beads, gel templating beads, or a combination thereof;
- gelating the fluidic droplets to form a plurality of gel beads using the solid templating beads, the gel templating beads, or a combination thereof,
- wherein the gel beads comprises the species and the solid templating beads, the gel templating beads, or a combination thereof.
The method of any preceding claim, further comprising:
- suspending the first gel beads in a third fluid sample comprising a plurality of second species;
- and
- reacting the one or more first species that have one or more chemical functionalities with one or more of the plurality of second species to functionalize the first gel beads with the one or more second species.
The method of any preceding claim, wherein the one or more first species that have one or more chemical functionalities are not reactive with the other first species.
The method of any preceding claim, wherein the chemical functionality is amino, ammonium, hydroxyl, mercapto, sulfone, sulfinic acid, sulfonic acid, thiocyanate, thione, thial, thiol, carboxyl, halocarboxy, halogen, imido, anhydrido, alkenyl, strained alkene, strained cycloalkene, alkynyl, strained alkyne, strained cycloalkyne, phenyl, benzyl, carbonyl, formyl, haloformyl, carbonato, ester, alkoxy, phenoxy, hydroperoxy, peroxy, ether, glycidyl, epoxy, hemiacetal, hemiketal, acetal, ketal, orthoester, orthocarbonate ester, amido, imino, imido, azido, azo, cyano, nitrato, nitrilo, nitrito, nitro, nitroso, pyridinyl, phosphinyl, phosphonic acid, phosphate, phosphoester, phosphodiester, boronic acid, boronic ester, borinic acid, borinic ester, acrylate, methacrylate, alkylacrylate, alkyne, terminal alkyne, nitrone, biotin, streptavidin, dibenzocyclooctyl, cholesterol, trans-cyclooctene, tetrazine, any derivatives thereof, or any combinations thereof.
The method of any preceding claim, wherein at least one of the first species is magnetic.
The method of any preceding claim, wherein at least one of the first species is colored and designed to color the first gel beads.
The method of any preceding claim, wherein at least part of the gelation is reversible.
The method of any preceding claim, further comprising dissolving the first gel beads.
The method of any preceding claim, further comprising disrupting at least one of first species to dissolve the first gel beads.
The method of any preceding claim, wherein one or more of the first species are second gel beads.
The method of any preceding claim, wherein the plurality of first species comprise one or more monomers comprising a first chemical functionality that can facilitate crosslinking of the one or more monomers.
The method of any preceding claim, wherein the first chemical functionality comprises one or more of methacrylate, methacrylamide, acrylate, methacrylate, alkylacrylate, acrylamide, methacrylamide, alkylacrylamide, alkylmethacrylamide, strained alkene, strained cycloalkene, alkynyl, strained alkyne, strained cycloalkyne, epoxy, azido, dibenzocyclooctyl, or derivatives thereof.
The method of any preceding claim, wherein the plurality of first species comprise at least one monomer comprising a second chemical functionality that does not facilitate crosslinking of the one or more monomers.
The method of any preceding claim, wherein the second chemical functionality comprises one or more of amino, hydroxyl, mercapto, sulfone, thiol, halogen, strained alkene, strained cycloalkane, dibenzocyclooctyl, epoxy, hemiacetal, hemiketal, acetal, ketal, amido, imino, imido, carboxylic acid, acyl halide, or azido.
The method of any preceding claim, wherein the plurality of second species comprise at least one oligomer comprising a third chemical functionality that is configured to react with the second chemical functionality.
The method of any preceding claim, wherein the third chemical functionality comprises one or more of amino, hydroxyl, mercapto, sulfone, thiol, halogen, strained alkene, strained cycloalkane, dibenzocyclooctyl, epoxy, hemiacetal, hemiketal, acetal, ketal, amido, imino, imido, carboxylic acid, acyl halide, or azido.
The method of any preceding claim, wherein the at least one oligomer comprise nucleotides, locked nucleotides, proteins, peptides, or peptide nucleic acids.
The method of any preceding claim, wherein the gel beads are surface-functionalized with a second chemical functionality.
The method of any preceding claim, wherein the gel beads comprise polyacrylamide.
The method of any preceding claim, wherein the gel beads are surface-functionalized with an amino group.
The method of any preceding claim, wherein a total number of oligomers functionalized on the surface of the gel beads is at least 90% higher than a total number of oligomers functionalized to the bulk of the gel beads.
The method of any preceding claim, wherein the plurality of species comprises a reagent that leads to gelation.
The method of any preceding claim, wherein the plurality of species comprises a biological.
The method of any preceding claim, wherein the gel templating beads are surface functionalized with a chemical functionality that is configured to react with the species.
Dependencies are inferred from the claim text.