SiM20 Posters

5 Sensors in Medicine 2020 • Monoliths well polymerized and anchored to the microfluidic channel walls • Pores randomly distributed across the monolith ATTACHMENT OFANTIBODY TO THE GMAMONOLITH • Antibodies were immobilized on the epoxy groups on GMA in monoliths • Solution (4 µL) added to the reservoir and flowed through the monolith by capillary action • After adding antibody, reservoirs filled with buffer and sealed to prevent evaporation • Reaction overnight at room temperature to provide sufficient time for primary amines on the antibody to react with epoxy groups on the GMA monolith • Remaining epoxy groups in the monolith blocked by flowing 0.1 M Tris buffer pH 8.5 through the monolith for 1 h Figure 3. Device operation and labeled antibody attachment. (A) Diagram of device operation for extraction of PTB biomarkers. The labeled PTB biomarker is flowed through the channel by either applying voltage or pressure. As labeled analyte passes the detection point, the signal is recorded. (B-C) CCD images of control (no anti-CRF flowed) and labeled anti-CRF attached to an immunoaffinity monolith. (D) Background subtracted fluorescence of monolith after reaction with labeled anti- CRF. PTB BIOMARKER EXTRACTION Figure 4. Fluorescence elution profile of 300 nM AF532-labeled CRF on an anti-CRF monolith or a control monolith lacking attached antibody from (A) buffer and (B) human blood serum (diluted 5-fold). CONCLUSIONS • Use of affinity monoliths in 3D printed microfluidic devices • GMA monolith well polymerized and anchored to the microfluidic channel walls • Captured PTB protein and peptide biomarkers on an affinity monolith in a 3D printed microfluidic device. FUTURE WORK • Create affinity monoliths with antibodies to all 9 PTB biomarkers • Extract PTB biomarkers from blood serum in a 3D printed microfluidic device Figure 2. SEM images of monoliths prepared from GMA in a 3D printed microfluidic device. (A) Channel view and (B) zoom view. Figure 5. Fluorescence elution profile of 300 nM AF532- labeled TNF on an anti-TNF monolith or a control monolith lacking attached antibody from (A) buffer and (B) human blood serum (diluted 20-fold).