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Methods for capturing nascent proteins

  • US 20100075374A1
  • Filed: 10/31/2008
  • Published: 03/25/2010
  • Est. Priority Date: 11/08/2007
  • Status: Abandoned Application
First Claim
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20. A method of generating and capturing nascent proteins, comprising:

  • (a) providing i) nucleic acid encoding a protein or fragment thereof, ii) a plurality of beads, each bead comprising one or more amplification primers and one or more first binding molecules, and iii) a population of second binding molecules capable of binding to said protein and said first binding molecules;

    (b) contacting said beads with said nucleic acid under conditions such that at least a portion of said nucleic acid is amplified to create treated beads comprising immobilized amplified nucleic acid;

    (c) contacting said treated beads with said second binding molecules under conditions such that at least a portion of said first binding molecules bind to at least a portion of said second binding molecules so as to create capture beads; and

    (d) producing nascent protein or fragments thereof from at least a portion of said immobilized amplified nucleic acid on said capture beads by cell free expression, at least a portion of said nascent protein or fragments thereof interacting with at least a portion of said second binding molecules so as to generate loaded beads comprising captured nascent protein or captured fragments thereof.

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