On-line aptamer affinity solid-phase extraction direct mass spectrometry for the rapid analysis of ?-synuclein in blood

On-line aptamer affinity solid-phase extraction direct mass spectrometry (AA-SPE-MS) is presented for the rapid purification, preconcentration, and characterization of alpha-synuclein (alpha-syn), which is a protein biomarker related to Parkinson's disease. Valve-free AA-SPE-MS is easily implemented using the typical SPE microcartridges and instrumental set-up necessary for on-line aptamer affinity solid-phase extraction capillary electrophoresis-mass spectrometry (AA-SPE-CE-MS). The essential requirement is substituting the application of the separation voltage by a pressure of 100 mbar for mobilization of the eluted protein through the capillary towards the mass spectrometer. Under optimized conditions with recombinant alpha-syn, repeatability is good in terms of migration time and peak area (percent relative standard deviation (%RSD) values (n = 3) are 1.3 and 6.6% at 1 mu g mL-1, respectively). The method is satisfactorily linear between 0.025 and 5 mu g mL-1 (R2 > 0.986), and limit of detection (LOD) is 0.02 mu g mL-1 (i.e. 1000, 500, and 10 times lower than by CE-MS, direct MS, and AA-SPE-CEMS, respectively). The established AA-SPE-MS method is further compared with AA-SPE-CE-MS, including for the analysis of alpha-syn in blood. The comparison discloses the advantages and disadvantages of AA-SPE-MS for the rapid and sensitive targeted analysis of protein biomarkers in biological fluids.

Automated summary

This study presents an online aptamer affinity solid-phase extraction direct mass spectrometry (AA-SPE-MS) method for the rapid purification, preconcentration, and characterization of alpha-synuclein (alpha-syn), a protein biomarker related to Parkinson's disease. The method utilizes SPE microcartridges and replaces the separation voltage with a pressure of 100 mbar to elute the protein towards the mass spectrometer. The developed method shows good repeatability, satisfactory linearity, and lower limits of detection compared to other techniques.