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US 2023/0416816 A1Published application
Methods and devices for mulitplexed proteomic and genetic analysis and on-device preparation of cdna
AssigneeIsoPlexis Corporation
InventorsRui ZHENG, Dana ASHWORTH, Sean MACKAY
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Abstract
Disclosed are devices and methods capable of multiplexed analysis of multiple cellular activities and pathways in single cells including genomic, transcriptomic, and proteomic analysis.
Claims38
A multiplex assay chip device configured for multiplexed analysis of biological material, the device comprising:
- a plurality of capture beads (CB) each bead including a capture moiety;
- and
- a substrate having a plurality of chambers each including
- an open end arranged on a first side of the substrate, and a length, a width, and a depth (“dimensions”),
- a pocket P1 having a pocket diameter greater than the width of the chamber, and
- at least one CB arranged within the pocket of each chamber,
- wherein each CB includes a CB diameter smaller than the pocket diameter and larger than the width of the chamber;
- and
- a surface configured to couple with the first side of the substrate to cover each chamber.
6 more elements
A multiplex assay chip device configured for multiplexed analysis of biological material, the device comprising:
- a plurality of capture beads (CB) each bead including a capture moiety;
- a substrate having a plurality of chambers each including
- an open end arranged on a first side of the substrate, and a length, a width, and a depth (“dimensions”),
- a plurality of pockets comprising at least a first pocket P1 and a second pocket P2, each pocket of the plurality of pockets having a pocket diameter greater than the width of the chamber, and
- at least one CB arranged within each pocket of each chamber, wherein each CB includes a CB diameter smaller than the pocket diameter and larger than the width of the chamber;
- and
- a surface configured to couple with the first side of the substrate to cover each chamber.
4 more elements
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A multiplexed method for the simultaneous identification of:
- (a) at least one nucleic acid sequence, and (b) at least one protein from a single subject cell comprising:
- (I) providing a device comprising:
- a substrate having a plurality of chambers each comprising at least one capture bead (CB), wherein a first CB, CB1, is configured to capture a target nucleic acid;
- and
- a surface configured to couple with a first side of the substrate to cover each chamber wherein:
- the surface comprises a plurality of substantially parallel lines of capture antibodies, each line of capture antibodies comprising a different antibody, configured to bind to a different target molecule, and at least one portion of each of the plurality of substantially parallel lines are arranged as to be exposed to each chamber;
- (II) introducing the subject cell to a chamber of the plurality of chambers;
- (III) maintaining the subject cell in the chamber under conditions sufficient to cause cell lysis to produce a cell lysate;
- (IV) identifying at least one first nucleic acid sequence, comprising:
- (a) providing conditions sufficient for the target nucleic acid to contact the CB1 to form a CB1-target nucleic acid complex,
- (b) determining the sequence of the complexed target nucleic acid to determine the sequence of the at least one expressed target nucleic acid sequence
- (V) detecting the at least one protein in the subject cell comprising the steps of:
- (a) incubating the cell lysate in the chamber under conditions sufficient to allow at least one antibody and at least one protein to form an antibody:protein complex;
- and
- (b) imaging the surface comprising the at least one antibody:protein complex, thereby identifying the one or more protein of the subject cell.
12 more elements
-134. (canceled)
Dependencies are inferred from the claim text.