Spatially encoded biological assays
First Claim
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1. A method for determining a location of a polypeptide and a nucleic acid in a biological sample, the method comprising:
- (a) contacting the biological sample with a binding agent that binds specifically to the polypeptide, wherein the binding agent is attached to a coding identifier having an oligonucleotide sequence;
(b) contacting the biological sample with an array comprising a first encoded probe and a second encoded probe, wherein;
the first encoded probe comprises (i) a binding agent that binds specifically to the oligonucleotide sequence of the coding identifier, and (ii) a coding tag comprising an oligonucleotide sequence that corresponds to a location in the array; and
the second encoded probe comprises (i) a binding agent that binds specifically to the nucleic acid, and (ii) a coding tag comprising an oligonucleotide sequence that corresponds to a location in the array;
(c) determining (i) all or a portion of the oligonucleotide sequence of the coding identifier, or a complement thereof, and (ii) all or a portion of the oligonucleotide sequence of the coding tag of the first encoded probe, or a complement thereof, and using the determined sequences of (i) and (ii) to determine the location of the polypeptide in the biological sample; and
(d) determining (i) all or a portion of the sequence of the nucleic acid, or a complement thereof, and (ii) all or a portion of the oligonucleotide sequence of the coding tag of the second encoded probe, or a complement thereof, and using the determined sequences of (i) and (ii) to determine the location of the nucleic acid in the biological sample.
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Abstract
The present invention provides assays and assay systems for use in spatially encoded biological assays. The invention provides an assay system comprising an assay capable of high levels of multiplexing where reagents are provided to a biological sample in defined spatial patterns; instrumentation capable of controlled delivery of reagents according to the spatial patterns; and a decoding scheme providing a readout that is digital in nature.
308 Citations
30 Claims
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1. A method for determining a location of a polypeptide and a nucleic acid in a biological sample, the method comprising:
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(a) contacting the biological sample with a binding agent that binds specifically to the polypeptide, wherein the binding agent is attached to a coding identifier having an oligonucleotide sequence; (b) contacting the biological sample with an array comprising a first encoded probe and a second encoded probe, wherein; the first encoded probe comprises (i) a binding agent that binds specifically to the oligonucleotide sequence of the coding identifier, and (ii) a coding tag comprising an oligonucleotide sequence that corresponds to a location in the array; and the second encoded probe comprises (i) a binding agent that binds specifically to the nucleic acid, and (ii) a coding tag comprising an oligonucleotide sequence that corresponds to a location in the array; (c) determining (i) all or a portion of the oligonucleotide sequence of the coding identifier, or a complement thereof, and (ii) all or a portion of the oligonucleotide sequence of the coding tag of the first encoded probe, or a complement thereof, and using the determined sequences of (i) and (ii) to determine the location of the polypeptide in the biological sample; and (d) determining (i) all or a portion of the sequence of the nucleic acid, or a complement thereof, and (ii) all or a portion of the oligonucleotide sequence of the coding tag of the second encoded probe, or a complement thereof, and using the determined sequences of (i) and (ii) to determine the location of the nucleic acid in the biological sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification