Direct clone analysis and selection technology

Abstract

The present invention describes a spatial addressing technique that uses a very high-density micro-pore array for high-throughput screening of biological interactions. The therapeutic, diagnostic, and drug-discovery implications of being able to identify, select and characterize specific protein-protein, protein-DNA and/or protein-carbohydrate interactions from heterogeneous populations of millions (to billions) of cells is discussed. Importantly, this technique possesses the screening and selection capacity of current display-based screening systems (i.e. millions-billions) but with greater efficiency and shorter time.

1 . A device comprising an array of micro-pores, the micro-pore array being reversibly attached to a solid substrate, wherein at least one binding partner is attached to said solid substrate, and wherein the internal diameter of the micro-pores ranges between approximately 1.0 micrometers and 500 micrometers. 
     
     
         2 . The device of  claim 1 , wherein the micro-pores are not coated with at least one binding partner. 
     
     
         3 - 4 . (canceled) 
     
     
         5 . The device of  claim 1 , wherein said device further comprises a polymeric film, wherein said array is covered by said polymeric film, wherein said polymeric film further comprises at least one hole and wherein said hole is positioned over at least one of said micro-pores. 
     
     
         6 . The device of  claim 5 , wherein said device further comprises a pressure source or electrolytic expulsion source configured proximal to said at least one hole. 
     
     
         7 . (canceled) 
     
     
         8 . The device of  claim 1 , wherein said micro-pores further comprise at least one biological cell. 
     
     
         9 . (canceled) 
     
     
         10 . The device of  claim 1 , wherein said device comprises a micro-pore testbed array, said array comprising a plurality of longitudinally fused fibers reversibly bonded to a single gas permeable solid substrate, wherein said solid substrate is degassed and wherein said solid substrate is attached to at least one binding partner. 
     
     
         11 . (canceled) 
     
     
         12 . The device of  claim 10 , comprising between approximately 300 to 11,500,000 of said fused fibers, per cm 2  of the array. 
     
     
         13 . The device of  claim 1 , wherein said solid substrate comprises a gas permeable material. 
     
     
         14 . The device of  claim 10 , wherein said gas permeable material comprises poly(dimethylsiloxane) (PDMS). 
     
     
         15 . The device of  claim 10  comprising said plurality of longitudinally fused fibers and a polymeric film, wherein said fused fibers are covered by said polymeric film, wherein said polymeric film further comprises at least one hole and wherein said hole is positioned over at least one of said fused fibers. 
     
     
         16 . The device of  claim 15 , wherein said device further comprises a pressure source configured proximal to said at least one hole. 
     
     
         17 .- 18 . (canceled) 
     
     
         19 . A method for identifying a sub-population of cells from a heterologous population of biological cells, the method comprising:
 a) providing:
 i) an array of micro-pores, wherein the internal diameter of micro-pores ranges between approximately 1.0 micrometers and 500 micrometers; 
 ii) said heterologous population of cells; 
 iii) at least one binding partner; 
   b) contacting said array with said heterologous population of cells and said at least one binding partner such that a sub-population comprising at least one of said biological cells settles into at least one of said micro-pores of said array;   c) incubating said array under conditions to promote the secretion of molecules from said biological cells; and   d) detecting desired secreted molecules in at least one of said micro-pores of said array, thereby identifying said sub-population of cells.   
     
     
         20 . The method of  claim 19 , wherein the providing step comprises providing an array of micro-pores not being coated with at least one binding partner. 
     
     
         21 - 45 . (canceled)