Berlin 2018 – wissenschaftliches Programm

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CPP: Fachverband Chemische Physik und Polymerphysik

CPP 37: Focus: Smart Hydrogels and Hydrogel Based Devices II - organized by Gerald Gerlach, Walter Richtering and Thomas Hellweg

CPP 37.4: Vortrag

Mittwoch, 14. März 2018, 10:30–10:45, C 130

Gene expression in NTA-hyaluronan hydrogel particles: A strategy for in situ separation and purification of His-tagged proteins — •THOMAS HEIDA, TONY KÖHLER, ANDREAS FERY, and JULIAN THIELE — Leibniz Institut für Polymerforschung e.V., Dresden, Germany

We present the synthesis and characterization of nitrilotriacetic acid (NTA)-modified hydrogel particles as well as their application in in vitro transcription/translation (IVTT), to selectively catch and separate expressed proteins in situ. Droplet microfluidics is employed to fabricate hydrogel particles based on furan-functionalized hyaluronic acid and maleimide-modified poly(ethylene glycol) (PEG) crosslinkers. As a proof of principle, we show selective conjugation and release of His-tagged green fluorescent protein (GFP) within the particles by reversible binding via Ni(II)-complexation. Thereafter, we utilize these particles as experimental platform in in vitro transcription/translation (IVTT) by additional functionalization with DNA encoding for His-GFP. In situ formed proteins are selectively separated from the complex IVTT reaction mixture and controllably released from the gel matrix after purification. Our NTA-hyaluronan microgels serve as robust experimental platform for cell-free synthesis and purification of His-tagged proteins.