Method and device for protein delivery into cells

• US 7,105,347 B2
• Filed: 07/30/2002
• Issued: 09/12/2006
• Est. Priority Date: 07/30/2002
• Status: Expired due to Fees

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Webb, Brian L., Fang, Ye, Lai, Fang, Picard, Laurent A. G.
• Primary Examiner(s): Guzo, David
• Assistant Examiner(s): Nguyen, Quang
• Application Number:
  US10/208,894
• Publication Number:
  US 20040023391A1
• Time in Patent Office: 1,505 Days
• Field of Search: 435/455, 435/320.1, 435/4, 435/7.2
• US Class Current: 435/455
• CPC Class Codes: C12N 2503/00 Use of cells in diagnostics
• CPC Class Codes: C12N 2535/10 Patterned coating
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

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.

Claims

1. A method of delivering a protein-of-interest into eukaryotic cells of primary cell lines, the method comprises:

providing a first population of cells that is easy to transfect relative to primary cells;

introducing into said first population of cells a DNA-construct encoding 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 secreted by said first population of cells;

capturing said protein-of-interest on said surface;

reacting said protein-of-interest with a carrier reagent;

depositing eukaryotic cells of primary cell lines over said protein-of-interest; and

, thereby said protein-of-interest is transported into eukaryotic cells of said primary cell lines.