Method and device for protein delivery into cells
First Claim
1. A method for performing surface-mediated protein delivery into living cells, the method comprising:
- providing a protein-containing mixture;
depositing said protein-containing mixture onto a surface at defined locations;
affixing said protein-containing mixture to the surface; and
plating said cells onto the surface in sufficient density and under conditions for delivering proteins into said cells.
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Abstract
Methods for performing surface-mediated protein delivery into living cells, and fabricating protein-transfected cell cluster arrays are provided. The method comprises providing a protein-containing mixture; depositing said protein-containing mixture onto a surface at defined locations; affixing the protein-containing mixture to the surface as microspots; and plating cells onto the surface in sufficient density and under conditions for the proteins to be delivered into the cells. The protein-containing mixture comprises any suitable amino acid sequence, including peptides, proteins, protein-domains, antibodies, or protein-nucleic acid conjugates, etc., with a carrier reagent. Protein-transfected cell arrays may be used for rapid and direct, screening of protein or enzymatic functions or any given intracellular protein interaction in the natural environment of a living cell, as well as for high-throughput screening of other biological and chemical analytes, which affect the functions of these proteins.
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Citations
104 Claims
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1. A method for performing surface-mediated protein delivery into living cells, the method comprising:
- providing a protein-containing mixture;
depositing said protein-containing mixture onto a surface at defined locations;
affixing said protein-containing mixture to the surface; and
plating said cells onto the surface in sufficient density and under conditions for delivering proteins into said cells. - 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)
- providing a protein-containing mixture;
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27. A reverse delivery method of introducing proteins into living cells comprising:
- spotting a protein of interest on a surface in defined and discrete locations;
drying and affixing the protein spot;
treating the protein spot with a carrier reagent;
depositing cells onto the surface; and
transporting said protein into said cells. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48)
- spotting a protein of interest on a surface in defined and discrete locations;
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49. A method of producing an array of cells transfected with a protein on a surface, the method comprising:
- spotting a protein-containing mixture onto a surface in discrete, defined locations;
drying said surface bearing the protein-containing mixture such that the spots remain affixed to the surface under array conditions;
adding cells to produce a surface having both proteins and cells; and
transporting of said proteins into said cells. - View Dependent Claims (50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60)
- spotting a protein-containing mixture onto a surface in discrete, defined locations;
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61. An array produced according to a method comprising:
- providing a protein-containing mixture;
depositing said protein-containing mixture onto a surface at defined locations;
affixing said protein-containing mixture to the surface; and
plating said cells onto the surface. - View Dependent Claims (62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103)
- providing a protein-containing mixture;
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85. A transfected-cell cluster array made according to a surface-mediated protein delivery method, the method comprising:
- providing DNA templates on a substrate surface;
synthesizing, in situ on said surface, proteins from said DNA templates;
affixing said proteins to said surface;
reacting said synthesized proteins with a carrier reagent;
adding cells to said surface; and
transporting said proteins into said cells.
- providing DNA templates on a substrate surface;
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104. A method of delivering a protein-of-interest into primary cell lines, the method comprises:
- providing a first population of cells;
introducing into said first population of cells a DNA-construct of said protein-of interest;
despositing said first population of cells onto a surface;
either lysing in situ said first population of cells, or allowing said protein-of-interest to be excreted by said first population of cells;
capturing said protein-of-interest on said surface;
reacting said protein-of-interest with a carrier reagent;
depositing a second population of cells over said protein-of-interest; and
, transporting said protein-of-interest into said second population of cells.
- providing a first population of cells;
Specification