TY - CONF A1 - Burnage, Samual Charles A1 - Bell, Jérémy A1 - Gawlitza, Kornelia A1 - Wan, Wei A1 - Rurack, Knut T1 - Microfluidic Platform for Functionalisation, Extraction and Detection of Phosphorylated Amino Acids Using Fluorescent Sensory Particles N2 - The reliable identification and quantification of phosphorylated amino acids, peptides and proteins is one of the key challenges in contemporary bioanalytical research, noteworthy, to diagnose and treat diseases at an early developmental stage. Miniaturised sensing devices like microfluidic chips combined with “smart” detection chemistry, simple data assessment, processing and presentation are very attractive for benchtop use in clinical environments. We developed novel synthetic probes targeting phosphorylated amino acids, based on core-shell microparticles consisting of a silica core coated with a molecularly imprinted polymer (MIP) shell. These “plastic antibodies” are extremely robust, resist denaturing solvents and elevated temperatures, can be reproducibly produced at low cost, and potentially overcome many of the practical problems in current bioanalytical detection strategies. The MIP layer contains a fluorescent probe monomer, binds selectively to phosphorylated tyrosine (pTyr) with a significant imprinting factor higher than 3.5 and responds with a “lighting-up” of its fluorescence accompanied by the development of a strongly red-shifted emission band toward the analyte. In analogy to our previous work [4], the bead-based ratiometric detection scheme has also been successfully transferred to a microfluidic chip format to demonstrate its applicability to rapid assays. Such a miniaturised device could yield an automated pTyr measurement system in the future. The setup was built by coupling a modular microfluidic system [5] for amino acid functionalisation (Fmoc protection) and a multi-layer PDMS/Teflon/glass microfluidic chip [6] for buffering, extraction (micropillars co-flow extraction) and selective adsorption on the MIP core-shell particles. A miniaturised optical assembly for low-light fluorescence measurements was also developed. Based on small opto-electronic parts and optical fibres, the emission from the MIP particles upon addition of pTyr concentrations from 0.5 – 200 μM could be monitored in real-time. T2 - IMA 2019 CY - Ioaninna, Greece DA - 22.09.2019 KW - MIPs KW - Microfluidics KW - Fluorescence KW - Sensing PY - 2019 AN - OPUS4-49201 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -