An injectable alginate-based hydrogel for microfluidic applications

•A new method for preparing injectable alginate monolith within microreactors is presented.•Gelation control was achieved in alginate based monolith by adjusting GDL concentration.•Stability of immobilized enzymes improved with chitosan addition.•High enzymatic conversion was observed in microreactor with developed immobilization technique. The objective of this study was to develop an injectable alginate based formulation for immobilizing enzymes into microfluidic systems. The gelation was induced upon lowering the pH by addition of d-glucono-δ-lactone (GDL) and release of Ca+ ions from solid CaCO3. The effects of GDL concentration on enzymatic activity and gelation time were investigated. The results indicated that increasing the GDL concentration increased both surface area and enzymatic activity. Also, chitosan was added to the formulation at different ratios to enhance the stability of enzyme during immobilization. For microfluidic application, 100μl spiral coil single channel microchip was fabricated and alginate GDL mixture containing β-glucosidase was injected to the microchannel prior to gelation. Enzymatic conversion was performed by pumping substrate (pNPG) through the microchannel. The results indicated that the entire substrate was converted continuously during 24h without any leakage or deactivation of immobilized enzyme.