Microchip-based homogeneous immunoassay using a cloned enzyme donor.
We have realized a cloned enzyme donor immunoassay (CEDIA) on a microchip in 96 s. CEDIA is a homogeneous immunoassay, based on the bacterial enzyme beta-galactosidase, which was genetically engineered into two inactive fragments: an enzyme donor and an enzyme acceptor. A model analyte was theophylline, and the detectable concentration range was from 0 to 40 microg mL(-1). Our CEDIA using a microfluidic device was very simple and rapid, unlike microchip-based heterogeneous immunoassays and CEDIA on a well-type microchip.
Tachi
Tomoya
T
Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Nagoya 464-8603, Japan.
Kaji
Noritada
N
Tokeshi
Manabu
M
Baba
Yoshinobu
Y
D001419:Bacteria / Q000201:enzymology; D005453:Fluorescence; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D046208:Microchip Analytical Procedures / Q000379:methods*; D046210:Microfluidic Analytical Techniques; D015202:Protein Engineering; D012680:Sensitivity and Specificity; D013806:Theophylline / Q000032:analysis / Q000276:immunology; D013997:Time Factors; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D013806:Theophylline; D001616:beta-Galactosidase