Jpn. J. Appl. Phys. 52 (2013) 06GK09 (5 pages)  |Previous Article| |Next Article|  |Table of Contents|
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Simultaneous Synthesis and Biotinylation of Proteins Using Puromycin-Based Labeling Technology for Fabrication of Protein Array Chip

Subhashini Raj Kumal1, Manish Biyani1,2, Shingo Ueno1,2, Takanori Akagi1, and Takanori Ichiki1,2

1Department of Bioengineering, School of Engineering, The University of Tokyo, Bunkyo, Tokyo 113-8656, Japan
2CREST, Japan Science and Technology Agency, Chiyoda, Tokyo 102-0075, Japan

(Received December 1, 2012; accepted January 18, 2013; published online June 20, 2013)

Protein arrays represent a class of devices that are of growing importance in the field of proteomics. These arrays enable screening of a large amount of proteins in a short time and at a lower cost. Here we present a method to fabricate protein array using biotin-conjugated puromycin to simultaneously synthesize and label proteins followed by immobilization onto streptavidin-functionalized surface based on the noncovalent biotin-streptavidin interaction. This method demonstrates the fabrication of protein array based on cell-free transcription/translation system using unmodified DNA as a starting genetic material. As a consequence, the procedure of protein arraying has been greatly simplified over the conventional approaches that require tedious and multi-step reactions. Further, an integrated approach of micro reactor array technology makes this method very simple and robust for achieving high-density protein arrays.

DOI: 10.7567/JJAP.52.06GK09

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