System and method for visualization and digital analysis of protein and other macromolecule microarrays
First Claim
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1. A method for visualization and digital analysis of sample interactions with a biological array, comprising:
- allowing the sample to interact with the array, which comprises biological molecules immobilized on a solid substrate in a two-dimensional and addressable pattern;
contacting the array with a secondary detector molecule comprising an enzyme;
incubating the array with a developing agent comprising a substrate of the enzyme, such that the enzyme catalyzes a reaction wherein the substrate is converted to a detectable product;
digitizing an array image created by the detectable product by scanning the array on a digital scanner; and
analyzing the digital image.
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Abstract
A system and method are disclosed for the rapid, reproducible and inexpensive imaging and digital analysis of molecular interactions between ligands and proteins and/or nucleic acids immobilized on an addressable two-dimensional microarray.
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Citations
16 Claims
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1. A method for visualization and digital analysis of sample interactions with a biological array, comprising:
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allowing the sample to interact with the array, which comprises biological molecules immobilized on a solid substrate in a two-dimensional and addressable pattern;
contacting the array with a secondary detector molecule comprising an enzyme;
incubating the array with a developing agent comprising a substrate of the enzyme, such that the enzyme catalyzes a reaction wherein the substrate is converted to a detectable product;
digitizing an array image created by the detectable product by scanning the array on a digital scanner; and
analyzing the digital image. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for analysis of sample interactions with a protein microarray, comprising:
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allowing the sample to interact with the microarray, comprising a plurality of proteins immobilized on a solid substrate in a two-dimensional and addressable pattern, the solid substrate comprising a barcode for sample identification and a PVDF membrane adhered to a rigid support;
contacting the microarray with a secondary detector molecule comprising a selective binding moiety and an enzyme conjugated thereto;
washing the microarray to remove unbound secondary detector molecules;
incubating the array with a developing agent comprising a substrate of the enzyme, such that the enzyme catalyzes a reaction wherein the substrate is converted to a detectable product;
washing the microarray to terminate the reaction and remove unreacted developing agent;
scanning the microarray using a digital scanner to create a digital image of the microarray; and
analyzing the digital image which corresponds to the sample interactions with the protein microarray.
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15. A system for analysis of autoimmune diseases in humans, comprising:
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a microarray, comprising a plurality of autoimmune markers immobilized on a solid substrate in a two-dimensional and addressable pattern, the solid substrate comprising a PVDF membrane adhered to a rigid support;
a first reagent comprising anti-human IgG conjugated to an enzyme;
a second reagent comprising a developing agent comprising a substrate of the enzyme, wherein the first and second reagents react when combined to yield a colorometric change; and
a flatbed digital scanner.
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16. A system for analysis of sample interactions with a biological array, comprising:
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an array, comprising a plurality of biological molecules immobilized on substrate in a two-dimensional and addressable pattern, the substrate comprising a rigid support which has an opaque surface or has been modified to have an opaque surface, said surface being adapted to bind the plurality of biological molecules;
a flatbed scanner adapted to produce a digital image of the array; and
a software program adapted to quantify and analyze the digital image.
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Specification