Sensitive analysis of α-synuclein by nonlinear laser wave mixing coupled with capillary electrophoresis

Sensitive analysis of α-synuclein by nonlinear laser wave mixing coupled with capillary electrophoresis

Multi-photon nonlinear laser wave-mixing spectroscopy is a novel absorption-based technique that offers excellent detection sensitivity for biomedical applications, including early diagnosis and investigation of neurodegenerative diseases. α-Synuclein is linked to Parkinson's disease (PD), and...

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Bibliographic Details
Published in:Analytical biochemistry Vol. 500; pp. 51 - 59
Main Authors: Iwabuchi, Manna F., Hetu, Marcel M., Tong, William G.
Format: Journal Article
Language:English
Published: United States Elsevier Inc 01-05-2016
Subjects:
alpha-Synuclein - analysis
alpha-Synuclein - chemistry
Amino Acid Sequence
Electrophoresis
Electrophoresis, Capillary - methods
Molecular Weight
Nonlinear Dynamics
Parkinson's disease
Spectrophotometry, Ultraviolet
Ultrasensitive nonlinear spectroscopy
α-Synuclein
CGE
α-Synuclein
UV
CE
Parkinson's disease
SDS
LIF
PAGE
CHES
SNARE
CZE
STEM
FITC
S/N
PEG
PD
LB
CSF
Electrophoresis
Ultrasensitive nonlinear spectroscopy
DMF
ELISA
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Summary:Multi-photon nonlinear laser wave-mixing spectroscopy is a novel absorption-based technique that offers excellent detection sensitivity for biomedical applications, including early diagnosis and investigation of neurodegenerative diseases. α-Synuclein is linked to Parkinson's disease (PD), and characterization of its oligomers and quantification of the protein may contribute to understanding PD. The laser wave-mixing signal has a quadratic dependence on analyte concentration, and hence the technique is effective in monitoring small changes in concentration within biofluids. A wide variety of labels can be employed for laser wave-mixing detection due to its ability to detect both chromophores and fluorophores. In this investigation, two fluorophores and a chromophore are studied and used as labels for the detection of α-synuclein. Wave-mixing detection limits of PD-related protein conjugated with fluorescein isothiocyanate, QSY 35 acetic acid, succinimidyl ester, and Chromeo P503 were determined to be 1.4 × 10−13 M, 1.4 × 10−10 M, and 1.9 × 10−13 M, respectively. Based on the laser probe volume used, the corresponding mass detection limits were determined to be 1.1 × 10−23 mol, 1.1 × 10−20 mol, and 1.5 × 10−23 mol. This study also presents molecular-based separation and quantification of α-synuclein by laser wave mixing coupled with capillary electrophoresis. •Ultrasensitive detection of α-synuclein using optical absorption-based laser method.•Monitor small changes in α-synuclein concentration levels by sensitive laser wave-mixing detection.•Modification of Chromeo P503TM reaction to detect protein at trace concentration levels.•Examine α-synuclein conformation using laser wave mixing coupled with capillary gel electrophoresis.•Fast analysis time and minimum sample preparation.
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content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2016.01.010