Compositions and methods for the simultaneous genomic, transcriptomic and proteomic analysis of single cells
Inventors
NG, Colin • KAISER, Alaina • BETTINI, Emily • Paczkowski, Patrick • Mackay, Sean • LIU, Feimei • Srinivasan, Maithreyan
Assignees
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Publication Number
US-12139748-B2
Publication Date
2024-11-12
Expiration Date
Abstract
Disclosed are compositions and methods for the multiplexed analysis of one or more intracellular targets of a single cell. Exemplary compositions of the disclosure comprise a surface comprising a plurality of capture agents operatively-linked thereto, wherein each capture agent specifically binds to a distinct intracellular target and wherein the plurality of capture agents form a repeating pattern; a substrate comprising a plurality of chambers, wherein the substrate releasably couples with the surface and wherein each chamber of the plurality of chambers comprises at least one repeat of the repeating pattern of the plurality of capture agents of the surface; a coating composition comprising a cell lysis composition; and a linker composition comprising a functionalization component and an extension component.
Core Innovation
The disclosed invention provides a device for multiplexed analysis of intracellular targets of a single cell. The device includes a surface comprising a plurality of antibodies operatively-linked thereto, where each antibody is configured to specifically bind to a distinct intracellular target, and the plurality of antibodies form a repeating pattern on the surface.
The device further includes a substrate comprising a plurality of chambers, where each chamber includes a biocompatible coating and a size exclusion well configured to accommodate a single bead, with a bead disposed within the size exclusion well. The substrate is configured to releasably couple with the antibody-patterned surface such that each chamber aligns and is in fluid communication with at least one repeat of the repeating pattern.
Each bead is coupled to an oligonucleotide comprising a barcode sequence and a sequence configured to specifically bind to a nucleic acid sequence of a cell. The abstract/summary describes embodiments that support simultaneous transcriptomic readouts via bead barcodes and oligo capture with downstream sequencing elements including UMI and template switching (TSO) features.
Claims Coverage
1 inventive feature is covered by independent claim clm-00001.
Antibody repeating-pattern surface aligned with chambered beads for multiplexed intracellular analysis
A device for multiplexed analysis of intracellular targets of a single cell, comprising a surface having a plurality of antibodies operatively-linked thereto that specifically bind distinct intracellular targets and form a repeating pattern; and a substrate with a plurality of chambers, each chamber having a biocompatible coating, a size exclusion well configured to accommodate a single bead, and a bead disposed within the well, wherein the substrate releasably couples with the surface so each chamber aligns and is in fluid communication with at least one repeat of the repeating pattern, and wherein the bead is coupled to an oligonucleotide comprising a barcode sequence and a sequence configured to specifically bind to a nucleic acid sequence of a cell.
The claim coverage centers on an antibody repeating-pattern surface that releasably aligns with a multi-chamber substrate, where single beads in size exclusion wells carry barcode oligonucleotides including a nucleic-acid binding sequence for multiplexed single-cell intracellular target analysis.
Stated Advantages
Documented Applications
No documented applications found
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