Shapeable and underwater super-elastic cellulose nanofiber/alginate cryogels by freezing-induced oxa-Michael reaction for efficient protein purification.

Construction of monolithic cryogels that can efficiently adsorb proteins is of great significance in biotechnological and pharmaceutical industries. Herein, a novel approach is presented to fabricate microfibrillated cellulose (MFC)/sodium alginate (SA) cryogels by using freezing-induced oxa-Michael reaction at -12 °C. Thanks to the controllable reactiveness of divinyl sulfone (DVS), cryo-concentrated pH increase activates the oxa-Michael reaction between DVS and hydroxyl groups of MFCs and SAs. The obtained composite cryogel exhibits outstanding underwater shape recovery and excellent fatigue resistance. Moreover, the MFC/SAs reveal a high lysozyme adsorption capacity of 294.12 mg/g, surpassing most of absorbent materials previously reported. Furthermore, the cryogel-packed column can purify lysozyme continuously from chicken egg white, highlighting its outstanding practical application performance. Reuse experiments indicated that over 90% of lysozyme extraction capacity was retained after 6 cycles. This work provides a new avenue to design and develop next-generation chromatographic media of natural polysaccharide-based cryogel for protein purification.