Highly sensitive microfluidic competitive enzyme immunoassay based on chemiluminescence resonance energy transfer for the detection of neuron-specific enolase.
A microfluidic 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 isothiocyanate-labeled 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.
Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, College of Chemistry and Pharmacy, Guangxi Normal University, Guilin, China.
Tingzhen
T
Yang
Marina
M
Vdovenko
Xue
X
Jin
Ivan Yu
IY
Sakharov
Shulin
S
Zhao
D016022:Case-Control Studies; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D057230:Limit of Detection; D016014:Linear Models; D008163:Luminescent Measurements / Q000379:methods*; D008175:Lung Neoplasms / Q000097:blood / Q000201:enzymology; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D010751:Phosphopyruvate Hydratase / Q000032:analysis* / Q000097:blood / Q000378:metabolism*; D015203:Reproducibility of Results
D010751:Phosphopyruvate Hydratase
Chemiluminescence; Enzyme immunoassays; Microchip electrophoresis; Neuron-specific enolase; Resonance energy transfer