Method and device for integrated protein expression, purification and detection
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United States of America Patent
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Issued Date -
Jul 31, 2003
app pub date -
Aug 22, 2002
filing date -
Aug 22, 2001
priority date (Note) -
Abandoned
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Abstract
A method for presenting a target protein or an array of target proteins for analysis is disclosed. The method includes the steps of forming a coding sequence; placing the coding sequence and protein-synthesis components capable of expressing the target protein under selected protein-synthesis conditions in a well on a substrate, the well having a surface which has been functionalized with the second coil-forming peptide; expressing said coding sequence under such conditions, wherein the target protein so synthesized binds to the well through coil-coil heterodimer formation, and is thus presented for analysis in the well in captured form; and washing said well to remove unbound components. The coding sequence includes a first nucleic acid sequence which encodes a first coil-forming peptide having a selected charge and being capable of interacting with a second, oppositely charged coil-forming peptide to form a stable .alpha.-helical coiled-coil heterodimer; and a second nucleic acid sequence encoding the target protein. Also disclosed are kits for use in practicing the method.
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