System, device and method for high-throughput multi-plexed detection
AssigneeYale University
InventorsRong Fan, Yao Lu, Jonathan Chen
Life of this patent
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Abstract
The present invention relates to a system, device, and method for the high throughput multiplexed detection of a wide number of compounds. The invention comprises of a microwell array coupled to a capture agent array to form a plurality of interfaces between a microwell and a set of immobilized capture agents. The set of capture agents comprises a plurality of distinguishable features, with each feature corresponding to the detection of a particular compound of interest. In certain embodiments, each microwell is configured to contain a single cell. The invention is therefore capable of performing a high throughput analysis of single cell profiles, including profiles of secreted compounds.
Claims17
A method for identifying proteins secreted from a single cell, the method comprising:
- (a) providing a device containing a plurality of enclosed interfaces, wherein each enclosed interface comprises a microchamber containing a cell and a substrate containing a plurality of spatially identifiable isolated features, wherein at least two of the isolated features contain at least two immobilized capture antibodies per isolated feature, and wherein each immobilized capture antibody recognizes a different protein of interest;
- (b) incubating the enclosed interfaces under conditions that result in secretion of proteins from cells and binding of the proteins secreted from the cells to capture antibodies immobilized on the isolated features of the substrate, thereby forming regions of the substrate that contain cell-secreted proteins captured on antibodies and correspond to the enclosed interfaces;
- (c) incubating the substrate of (b) in the presence of detection antibodies under conditions that result in binding of detection antibodies to captured proteins, wherein each detection antibody recognizes a different captured protein, wherein at least two of the isolated features are associated with at least two detection antibodies per isolated feature, and wherein each of the detection antibodies in an isolated feature contains a spectrally-distinct label relative to all other detection antibodies in the same isolated feature;
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- (d) detecting detection antibodies bound to captured proteins in a region of the substrate that corresponds to an enclosed interface of (a), thereby identifying proteins secreted from a cell contained within the corresponding enclosed interface.
A method for identifying proteins secreted from a single cell, the method comprising:
- (a) providing a device containing a plurality of enclosed interfaces, wherein each enclosed interface comprises a microchamber containing a cell and a substrate containing at least ten isolated features, wherein at least ten spatially identifiable of the isolated features contains at least two immobilized capture antibodies per isolated feature, and wherein each immobilized capture antibody recognizes a different protein of interest;
- (b) incubating the enclosed interfaces under conditions that result in secretion of proteins from cells and binding of the proteins secreted from the cells to capture antibodies immobilized on the isolated features of the substrate, thereby forming regions of the substrate that contain cell-secreted proteins captured on antibodies and correspond to the enclosed interfaces;
- (c) incubating the substrate of (b) in the presence of detection antibodies under conditions that result in binding of detection antibodies to captured proteins, wherein each detection antibody recognizes a different captured protein, wherein at least ten of the isolated feature are associated with at least two detection antibodies per isolated feature, and wherein each of the detection antibodies in an isolated feature contains a spectrally-distinct label relative to all other detection antibodies in the same isolated feature;
- and
- (d) detecting detection antibodies bound to captured proteins in a region of the substrate that corresponds to an enclosed interface of (a), thereby identifying proteins secreted from a cell contained within the corresponding enclosed interface.
Dependencies are inferred from the claim text.