Document Detail


Multiplexed amino acid array utilizing bioluminescent Escherichia coli auxotrophs.
MedLine Citation:
PMID:  20405822     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Moon Il Kim; Byung Jo Yu; Min-Ah Woo; Daeyeon Cho; Jonathan S Dordick; June Hyoung Cho; Byung-Ok Choi; Hyun Gyu Park
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  82     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-05-14     Completed Date:  2010-09-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4072-7     Citation Subset:  IM    
Affiliation:
Department of Chemical and Biomolecular Engineering, KAIST 335 Gwahak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis*
Escherichia coli / chemistry*
Humans
Infant
Infant, Newborn
Luminescence / diagnostic use*
Signal Transduction / physiology*
Chemical
Reg. No./Substance:
0/Amino Acids

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