Development of a NanoBioAnalytical platform for "on-chip" qualification and quantification of platelet-derived microparticles.
Blood microparticles (MPs) are small membrane vesicles (50-1000nm), derived from different cell types. They are known to play important roles in various biological processes and also recognized as potential biomarkers of various health disorders. Different methods are currently used for the detection and characterization of MPs, but none of these methods is capable to quantify and qualify total MPs at the same time, hence, there is a need to develop a new approach for simultaneous detection, characterization and quantification of microparticles. Here we show the potential of surface plasmon resonance (SPR) method coupled to atomic force microscopy (AFM) to quantify and qualify platelet-derived microparticles (PMPs), on the whole nano-to micro-meter scale. The different subpopulations of microparticles could be determined via their capture onto the surface using specific ligands. In order to verify the correlation between the capture level and the microparticles concentration in solution, two calibration standards were used: Virus-Like Particles (VLPs) and synthetic beads with a mean diameter of 53nm and 920nm respectively. The AFM analysis of the biochip surface allowed metrological analysis of captured PMPs and revealed that more than 95% of PMPs were smaller than 300nm. Our results suggest that our NanoBioAnalytical platform, combining SPR and AFM, is a suitable method for a sensitive, reproducible, label-free characterization and quantification of MPs over a wide concentration range (≈107to 1012particles/mL; with a limit of detection (LOD) in the lowest ng/µL range) which matches with their typical concentrations in blood.
FEMTO-ST Institute, UBFC, CNRS, ENSMM, UTBM, 15B Avenue des Montboucons, 25030 Besançon, France.
Sameh
S
Obeid
INSERM U1098, EFS Bourgogne Franche-Comté, UBFC, 25000 Besançon, France.
Adam
A
Ceroi
INSERM U1098, EFS Bourgogne Franche-Comté, UBFC, 25000 Besançon, France.
Guillaume
G
Mourey
INSERM U1098, EFS Bourgogne Franche-Comté, UBFC, 25000 Besançon, France.
Philippe
P
Saas
FEMTO-ST Institute, UBFC, CNRS, ENSMM, UTBM, 15B Avenue des Montboucons, 25030 Besançon, France.
Celine
C
Elie-Caille
FEMTO-ST Institute, UBFC, CNRS, ENSMM, UTBM, 15B Avenue des Montboucons, 25030 Besançon, France. Electronic address: wboireau@femto-st.fr.
Wilfrid
W
Boireau
D015374:Biosensing Techniques; D001792:Blood Platelets / Q000737:chemistry / Q000648:ultrastructure*; D055252:Cell-Derived Microparticles / Q000737:chemistry / Q000648:ultrastructure*; D005434:Flow Cytometry; D006801:Humans; D018625:Microscopy, Atomic Force; D020349:Surface Plasmon Resonance
Atomic force microscopy; Blood microparticles; Characterization; Platelets; Quantification; Surface plasmon resonance