AFFINITY-OLIGONUCLEOTIDE CONJUGATES AND USES THEREOF
First Claim
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1. A method for characterizing cells comprising:
- performing a reaction in at least one single vessel of a plurality of vessels, the reaction comprising attaching a first vessel barcoded polynucleotide, which comprises a first vessel barcode sequence, to an oligonucleotide portion of an affinity-oligonucleotide conjugate, wherein an affinity portion of the affinity-oligonucleotide conjugate binds to a target antigen expressed by a single cell isolated in the at least one single vessel of the plurality of vessels, and wherein the oligonucleotide portion comprises an antigen identification sequence (AID).
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Abstract
Provided herein are methods and composition for single cell characterization using affinity-oligonucleotide conjugates.
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Citations
124 Claims
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1. A method for characterizing cells comprising:
- performing a reaction in at least one single vessel of a plurality of vessels, the reaction comprising attaching a first vessel barcoded polynucleotide, which comprises a first vessel barcode sequence, to an oligonucleotide portion of an affinity-oligonucleotide conjugate, wherein an affinity portion of the affinity-oligonucleotide conjugate binds to a target antigen expressed by a single cell isolated in the at least one single vessel of the plurality of vessels, and wherein the oligonucleotide portion comprises an antigen identification sequence (AID).
- View Dependent Claims (6, 9, 18, 29, 30, 31, 41, 46, 56, 61, 64, 70, 73, 75, 79, 81, 84, 98, 106)
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2-5. -5. (canceled)
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7-8. -8. (canceled)
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10-17. -17. (canceled)
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19-28. -28. (canceled)
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32-40. -40. (canceled)
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42-45. -45. (canceled)
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47-55. -55. (canceled)
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57-60. -60. (canceled)
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62-63. -63. (canceled)
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65-69. -69. (canceled)
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71-72. -72. (canceled)
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74. (canceled)
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76-78. -78. (canceled)
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80. (canceled)
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82-83. -83. (canceled)
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85-97. -97. (canceled)
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99-105. -105. (canceled)
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107-113. -113. (canceled)
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114. A composition comprising a plurality of vessels, wherein at least one single vessel of the plurality of vessels comprises
a. a single cell from a sample comprising a plurality of cells, and b. a vessel barcoded polynucleotide or complement thereof comprising a vessel barcode sequence; - and
c. an affinity-oligonucleotide conjugate that binds to a target antigen of the single cell. - View Dependent Claims (120, 121)
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115-119. -119. (canceled)
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122. (canceled)
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123. A kit comprising:
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(a) a first container comprising a first oligonucleotide comprising a first antigen identification (AID) sequence, wherein the first AID sequence is a known sequence; (b) a second container comprising a second oligonucleotide comprising a second antigen identification (AID) sequence, wherein the second AID sequence is a known sequence and is different than the first AID sequence; (c) one or more third containers comprising reagents capable of conjugating the first oligonucleotide to a first affinity molecule and reagents capable of conjugating the second oligonucleotide to a second affinity molecule; (d) a set of instructions describing how to conjugate the first oligonucleotide to the first affinity molecule and the second oligonucleotide to the second affinity molecule.
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124. A kit comprising:
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(a) a first container comprising an oligonucleotide comprising an antigen identification (AID) sequence, wherein the AID sequence is a known sequence; (b) a second container comprising reagents capable of conjugating the oligonucleotide to an affinity molecule; (c) a third container comprising a plurality of vessel barcoded polynucleotides; and (d) a set of instructions describing how to attach a vessel barcoded polynucleotide of the plurality of vessel barcoded polynucleotides to the oligonucleotide when conjugated to the affinity molecule.
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Specification