NDLI: Integrated microfluidic enzyme reactor mass spectrometry platform for detection of anthrax lethal factor

Content Provider	IEEE Xplore Digital Library
Author	Aravamudhan, S. Joseph, P.J. Kuklenyik, Z. Boyer, A.E. Barr, J.R.
Copyright Year	2009
Description	Author affiliation: Centers for Disease Control and Prevention (CDC), National Center for Environmental Health, Division of Laboratory Sciences, Atlanta, GA 30341, USA (Kuklenyik, Z.; Boyer, A.E.; Barr, J.R.) \|\| Nanotechnology Research Center, Georgia Institute of Technology, Atlanta GA 30332 USA (Aravamudhan, S.; Joseph, P.J.)
Abstract	In this work, we have developed a coupled microfluidic enzyme reactor mass spectrometry platform for the detection of protein toxins such as anthrax lethal factor. The lethal toxin produced during Bacillus anthracis infection is a complex protective antigen, which localizes the toxin to the cell receptor and lethal factor (LF). We have demonstrated, in this work, the applicability of a microfluidic reactor for the capture and concentration of enzyme reaction solid-phase. The reaction solid-phase consists of anti-LF monoclonal antibodies immobilized on magnetic protein G beads for the capture of LF. The captured LF, on exposure to optimized peptide substrate, hydrolyzes into two smaller peptide products. These cleavage products were then analyzed by mass spectrometer coupled to the microfluidic reactor. This resulted in efficient sample preparation, high sensitivity, larger reaction sites, less reagents consumption and shorter analysis time. We have showed here reproducible detection of anthrax lethal factor in concentration range of 40 to 0.5 ng/mL with a detection limit of 1 ng/mL. The enzymatic reaction and the analysis were performed in less than 15 minutes, indicating a rapid diagnostic tool for early anthrax prognosis.
Starting Page	1071
Ending Page	1074
File Size	506850
Page Count	4
File Format	PDF
ISBN	9781424432967
ISSN	1557170X
DOI	10.1109/IEMBS.2009.5335064
Language	English
Publisher	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Publisher Date	2009-09-03
Publisher Place	USA
Access Restriction	Subscribed
Rights Holder	Institute of Electrical and Electronics Engineers, Inc. (IEEE)
Subject Keyword	Microfluidics Biochemistry Inductors Mass spectroscopy Proteins Peptides Diseases Protection Fluorescence Magnetic analysis
Content Type	Text
Resource Type	Article
Subject	Signal Processing Biomedical Engineering Health Informatics Computer Vision and Pattern Recognition

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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