On-line Aptamer Affinity Solid-Phase Extraction Capillary Electrophoresis-Mass Spectrometry for the Analysis of Blood α-Synuclein.
Roger Pero-GasconFernando BenaventeZoran MinicMaxim V BerezovskiVictoria Sanz-NebotPublished in: Analytical chemistry (2019)
In this paper, an on-line aptamer affinity solid-phase extraction capillary electrophoresis-mass spectrometry method is described for the purification, preconcentration, separation, and characterization of α-synuclein (α-syn) in blood at the intact protein level. A single-stranded DNA aptamer is used to bind with high affinity and selectivity α-syn, which is a major component of Lewy bodies, the typical aggregated protein deposits found in Parkinson's disease (PD). Under the conditions optimized with recombinant α-syn, repeatability (2.1 and 5.4% percent relative standard deviation for migration times and peak areas, respectively) and microcartridge lifetime (around 20 analyses/microcartridge) were good, the method was linear between 0.5 and 10 μg·mL-1, and limit of detection was 0.2 μg·mL-1 (100 times lower than by CE-MS, 20 μg·mL-1). The method was subsequently applied to the analysis of endogenous α-syn from red blood cells lysate of healthy controls and PD patients.
Keyphrases
- capillary electrophoresis
- mass spectrometry
- liquid chromatography
- solid phase extraction
- high performance liquid chromatography
- tandem mass spectrometry
- gas chromatography
- high resolution mass spectrometry
- liquid chromatography tandem mass spectrometry
- molecularly imprinted
- simultaneous determination
- ultra high performance liquid chromatography
- gas chromatography mass spectrometry
- gold nanoparticles
- label free
- sensitive detection
- red blood cell
- high resolution
- end stage renal disease
- ejection fraction
- binding protein
- cell free
- protein protein
- chronic kidney disease
- prognostic factors
- magnetic nanoparticles
- newly diagnosed
- peritoneal dialysis
- circulating tumor
- loop mediated isothermal amplification
- parkinson disease
- amino acid
- structural basis