Highly sensitive microfluidic competitive enzyme immunoassay based on chemiluminescence resonance energy transfer for the detection of neuron-specific enolaseстатья
Статья опубликована в высокорейтинговом журнале
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Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 2 сентября 2014 г.
Аннотация:Amicrofluidic competitive enzyme immunoassay based on chemiluminescence resonance
energy transfer (CRET) was developed for highly sensitive detection of neuron-specific enolase
(NSE). The CRET system consisted of horseradish peroxidase (HRP)/luminol as a light
donor and fluorescein isothiocyanate as an acceptor. When fluorescein isothiocyanatelabeled
antibody binds with HRP-labeled antigen to form immunocomplex, the donor
and acceptor are brought close each other and CRET occurs in the immunocomplex. In
the MCE, the immunocomplex and excess HRP–NSE were separated, and the chemiluminescense
intensity of immunocomplex was used to estimate NSE concentration.
The calibration curve showed a linearity in the range of NSE concentrations from 9.0 to
950 pM with a correlation coefficient of 0.9964. Based on a S/N of 3, the detection limit for
NSE determination was estimated to be 4.5 pM, which is two-order magnitude lower than
that of without CRET detection. This assay was applied for NSE quantification in human
serum. The obtained results demonstrated that the proposed immunoassay may serve as
an alternative tool for clinical analysis of NSE.