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Multiplexed amino acid array utilizing bioluminescent escherichia coli auxotrophs

Title
Multiplexed amino acid array utilizing bioluminescent escherichia coli auxotrophs
Authors
Kim M.I.Yu B.J.Woo M.-A.Cho D.Dordick J.S.Cho J.H.Choi B.-O.Park H.G.
Ewha Authors
최병옥
Issue Date
2010
Journal Title
Analytical Chemistry
ISSN
0003-2700JCR Link
Citation
vol. 82, no. 10, pp. 4072 - 4077
Indexed
SCI; SCIE; SCOPUS WOS scopus
Abstract
We describe a novel multiplex "amino acid array" for simultaneously quantifying different amino acids based on the rapid growth of amino acid auxotrophic E. coli. First, we constructed genetically engineered amino acid auxotrophs of E. coli containing a bioluminescence reporter gene, yielding concomitant luminescence as a response to cell growth, and then immobilized the reporter cells within individual agarose of respective wells in a 96-well plate serving as a mimic of a biochip. Using the amino acid array, we were able to determine quantitatively the concentrations of 16 amino acids in biological fluid by simply measuring bioluminescent signals from the immobilized cells within 4 h without pre- and post-treatment. The clinical utility of this method was verified by quantifying different amino acids in dried blood spot specimens from clinical samples for the diagnosis of metabolic diseases of newborn babies. This method serves as a convenient route to the rapid and simultaneous analysis of multiple amino acids from complex biological fluids and represents a new analytical paradigm that can replace conventional, yet laborious methods currently in use. © 2010 American Chemical Society.
DOI
10.1021/ac100087r
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의학전문대학원 > 의학과 > Journal papers
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