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Immuno-MALDI-MS in Human Plasma and On-Chip Biomarker Characterizations at the Femtomole Level
Institut FEMTO-ST, Université de Franche Comté, CLIPP (Clinical-Innovation Proteomic Platform), 25044 Besançon, France
Santé STIC, CLIPP (Clinical-Innovation Proteomic Platform), Université de Bourgogne, Centre Hospitalier Universitaire de Dijon, 21000 Dijon, France
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 8 October 2012; in revised form: 18 October 2012 / Accepted: 2 November 2012 / Published: 6 November 2012
(This article belongs to the Special Issue Biochips
Abstract: Immuno-SPR-MS is the combination of immuno-sensors in biochip format with mass spectrometry. This association of instrumentation allows the detection and the quantification of proteins of interest by SPR and their molecular characterization by additional MS analysis. However, two major bottlenecks must be overcome for a wide diffusion of the SPR-MS analytical platform: (i) To warrant all the potentialities of MS, an enzymatic digestion step must be developed taking into account the spot formats on the biochip and (ii) the biological relevancy of such an analytical solution requires that biosensing must be performed in complex media. In this study, we developed a procedure for the detection and the characterization at ~1 µg/mL of the LAG3 protein spiked in human plasma. The analytical performances of this new method was established, particularly its specificity (S/N > 9) and sensitivity (100% of LAG3 identification with high significant mascot score >68 at the femtomole level). The collective and automated on-chip MALDI-MS imaging and analysis based on peptidic fragments opens numerous applications in the fields of proteomics and diagnosis.
Keywords: Surface Plasmon Resonance; mass spectrometry; immuno MALDI-MS; biomarker; proteomics
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Rouleau, A.; Osta, M.E.; Lucchi, G.; Ducoroy, P.; Boireau, W. Immuno-MALDI-MS in Human Plasma and On-Chip Biomarker Characterizations at the Femtomole Level. Sensors 2012, 12, 15119-15132.
Rouleau A, Osta ME, Lucchi G, Ducoroy P, Boireau W. Immuno-MALDI-MS in Human Plasma and On-Chip Biomarker Characterizations at the Femtomole Level. Sensors. 2012; 12(11):15119-15132.
Rouleau, Alain; Osta, Marven E.; Lucchi, Géraldine; Ducoroy, Patrick; Boireau, Wilfrid. 2012. "Immuno-MALDI-MS in Human Plasma and On-Chip Biomarker Characterizations at the Femtomole Level." Sensors 12, no. 11: 15119-15132.