Improved specific biodetection with ion trap mobility spectrometry (ITMS): a 10-min, multiplexed, immunomagnetic ELISA.
Enabling trace chemical detection equipment utilized in the field to transduce a biodetection assay would be advantageous from a logistics, training, and maintenance standpoint. Described herein is an assay design that uses an unmodified, commercial off-the-shelf (COTS) ion trap mobility spectrometer to analyze an immunomagnetic enzyme-linked immunosorbant assay (ELISA). The assay, which uses undetectable enzymatic substrates and ELISA-generated detectable products, was optimized to quantitatively report the amount of target in the sample. Optimization of this ELISA design retained the assay specificity and detection limit (approximately 10(3) E. coli per assay) while decreasing the number of user steps and reducing the assay time to 10 min (>9-fold decrease as compared to past studies). Also discussed are previously undescribed, independent substrate/enzyme/product combinations used in the immunomagnetic ELISA. These discoveries allow for the possibility of a quantitative, multiplexed, 10-min assay that is analyzed by the ion trap mobility spectrometer trace chemical detector.
Pris
Andrew D
AD
General Electric-Global Research Center, Niskayuna, New York 12309, USA. pris@research.ge.com
Mondello
Frank J
FJ
Wroczynski
Ronald J
RJ
Murray
Anthony J
AJ
Boudries
Hacene
H
Surman
Cheryl M
CM
Paxon
Tracy L
TL
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005696:Galactosidases / Q000737:chemistry / Q000378:metabolism; D005697:Galactosides / Q000737:chemistry; D005966:Glucuronidase / Q000737:chemistry / Q000378:metabolism; D006912:Hydroxyquinolines / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D009596:Nitrophenols / Q000737:chemistry; D011732:Pyridoxal Phosphate / Q000737:chemistry
D005697:Galactosides; D006912:Hydroxyquinolines; D009596:Nitrophenols; C056147:8-hydroxyquinoline glucuronide; D011732:Pyridoxal Phosphate; D000469:Alkaline Phosphatase; D005696:Galactosidases; D005966:Glucuronidase