Protein arrays and methods of using and making the same

Abstract

Methods and devices are provided for preparing a protein array having a plurality of proteins. In one embodiment, the method includes providing a plurality of nucleic acids each having a predefined sequence and expressing in vitro a plurality of proteins from the plurality of nucleic acids. In another embodiment, protein arrays having a solid surface and a microvolume are also provided. The solid surface can have a plurality of anchor oligonucleotides capable of hybridizing with a plurality of nucleic acids. The microvolume can cover each of the plurality of anchor oligonucleotides and can be configured to produce a polypeptide from each of the plurality of nucleic acids.

What is claimed is;: 
     
         1 . A method for preparing a protein array having a plurality of proteins, the method comprising:
 (a) providing a plurality of nucleic acids each having a predefined sequence; and   (b) expressing in vitro a plurality of proteins from the plurality of nucleic acids, wherein the plurality of proteins are expressed on an array.   
     
     
         2 . The method of  claim 1  further comprising (c) measuring an activity of each of the plurality of proteins. 
     
     
         3 . The method of  claim 1 , wherein the plurality of nucleic acids are synthesized on a solid surface. 
     
     
         4 . The method of  claim 1 , wherein each of the plurality of nucleic acids comprises a regulatory genetic sequence. 
     
     
         5 . The method of  claim 1 , wherein each of the plurality of proteins is expressed in vitro in a well of a micro-well plate. 
     
     
         6 . The method of  claim 1 , wherein each of the plurality of proteins is expressed in vitro at a different feature of a solid surface. 
     
     
         7 . A method for preparing a protein array having a plurality of proteins, the method comprising:
 (a) providing a first microvolume comprising a population of nucleic acids having a plurality of distinct, predefined sequences;   (b) immobilizing the nucleic acid sequences onto an array comprising a plurality of anchor oligonucleotides having a sequence complementary to a terminus sequence of the nucleic acids;   (c) expressing in vitro in a second microvolume a plurality of proteins from the population of nucleic acids.   
     
     
         8 . A method for producing at least one protein, the method comprising:
 (a) providing a support having a plurality of distinct features, each feature comprising a plurality of immobilized anchor oligonucleotides;   (b) generating at least one plurality of nucleic acid having a predefined sequence onto the plurality of anchor oligonucleotides;   (c) providing a microvolume onto at least one feature of the support; and   (d) expressing in vitro in the microvolume the at least one protein from the at least one nucleic acid.   
     
     
         9 . The method of  claim 8  wherein each feature of the support comprises a distinct plurality of support-bound anchor oligonucleotides, wherein the 5′ end of each of the plurality of anchor oligonucleotide is complementary to the 5′ end of a distinct nucleic acid having a predefined sequence. 
     
     
         10 . The method of  claim 8  wherein the plurality of nucleic acids are generated by assembling a plurality of construction oligonucleotides comprising partially overlapping sequences that define the sequence of the at least one nucleic acid. 
     
     
         11 . The method of  claim 10  wherein the at least one nucleic acid is generated under (i) ligation conditions, (ii) chain extension conditions, or (iii) chain extension and ligation conditions. 
     
     
         12 . The method of  claim 8  wherein the microvolume comprises reagents appropriate for expressing in vitro the at least one protein from the at least one nucleic acid. 
     
     
         13 . The method of  claim 8  further comprising verifying the at least one nucleic acid sequence prior to the step of expressing the at least one protein. 
     
     
         14 . The method of  claim 8  further comprising:
 synthesizing a plurality of partially overlapping construction oligonucleotides, wherein each construction oligonucleotide is synthesized at a distinct feature of the support comprising immobilized complementary construction oligonucleotides; 
 releasing the construction oligonucleotides in at least one microvolume; and 
 transferring the at least one microvolume to a distinct feature comprising a plurality of anchor oligonucleotides. 
 
     
     
         15 . The method of  claim 8  wherein at least 1,000 proteins are produced. 
     
     
         16 . The method of  claim 8  wherein the proteins are proteins variants. 
     
     
         17 . The method of  claim 8  further comprising screening the at least one protein to identify proteins having a desired characteristic. 
     
     
         19 . A protein array comprising:
 a solid surface having a plurality of anchoroligonucleotides capable of hybridizing with a plurality of nucleic acids; and   a microvolume covering each of the plurality of anchor oligonucleotides, the microvolume configured to produce a polypeptide from each of the plurality of nucleic acids.   
     
     
         20 . A protein array comprising:
 (a) a first plurality of features on a support, each of the first plurality of features comprising a plurality of immobilized single stranded oligonucleotides, wherein the plurality of single stranded oligonucleotides comprises partially overlapping sequences that define the sequence of each of a plurality of nucleic acid molecules encoding a plurality of proteins; and   (b) a second plurality of features, the second plurality of features comprising a plurality of anchor oligonucleotides having a sequence complementary to a terminus sequence of each of the plurality nucleic acids.