In this presentation, two methods involving synthetic, water-soluble polymers for the stabilization of enzymes, and most generally proteins, will be discussed.<br/>On the one hand, end-functional polymers obtained by RAFT polymerization are assessed as alternative to PEGylation in a biotechnological/pharmaceutical context (Figure A). Straightforward conjugation experiments are followed by careful chromatographic purification leading to the isolation of semi-discrete protein-polymer species. The latter enable a more precise evaluation of the effect of conjugation on the stabilization of proteins, which is studied in various conditions to produce relevant phase diagrams. On the other hand, we investigate the coating of enzymes with a thin layer of hydrogel (Figure B). Particularly, we present a simplified protocol for single-protein encapsulation that maintains the nanoscopic character of the protein, an important feature for bionanotechnology, yet with a full coverage of the enzyme surface. This leads to enhanced resistance of the biocatalysts towards organic solvents or pH variations. Through an extensive study with a range of proteins with various functions and sizes, we postulate the universality of the method. The intrinsic reactivity of the hydrogel coating is further exploited for labeling and surface immobilization.

Semi-discrete protein-RAFT polymer conjugates and single-enzyme nanogels

2019 Spring National Meeting

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