On-the-fly rapid immunoassay for neonatal sepsis diagnosis: C-reactive protein accurate determination using magnetic graphene-based micromotors

Based on the exceptional and new opened biosensing possibilities of self-propelled micromotors, a micromotor-based immunoassay (MIm) has smartly been designed for C-reactive protein (CRP) determination in plasma of preterm infants with sepsis suspicion. The design of the micromotors involved the electrosynthesis of a carbon-based outer layer (for antibody functionalization), an intermediate Ni layer (for magnetic guidance and stopped flow operations) and PtNPs inner catalytic layer (for catalytic bubble propulsion). Micromotors biofunctionalization on the outer layer (using carbon black (CB), reduced graphene oxide (rGO) and multi-walled carbon nanotubes (MWCNTs), and biocompatible propulsion capabilities, were carefully studied. Magnetic rGO/Ni/PtNPs micromotors exhibited the most efficient and reproducible (CV = 9%) anti-CRP functionalization, controlled stopped-flow operations as well as efficient bubble propulsion (1% H2O2, 1,5% NaCh, speed 140 mu m s(-1)). Analytical performance of MIm was excellent, allowing the direct (without dilution), sensitive (LOD = 0.80 mu g/mL), and accurate CRP determination (E-r = 1%) in hardly available preterm babies' plasma samples with suspected sepsis using very low volumes (<10 mu L) and in just 5 min of on-the-fly bioassay. Overall, the results obtained allowed the fast and reliable sepsis diagnostics in preterm babies' individuals with suspected sepsis, not only proving the usefulness of the approach as its potential utilization as point-of-care device for clinical analysis but drawing new horizons in extremely low sample volumes-based diagnostics.