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Antifreeze proteins from polar fish species are remarkable biomacromolecules which prevent the growth of ice crystals. Ice crystal growth is a major problem in cell/tissue cryopreservation for transplantation, transfusion and basic biomedical research, as well as technological applications such as icing of aircraft wings. This review will introduce the rapidly emerging field of synthetic macromolecular (polymer) mimics of antifreeze proteins. Particular focus is placed on designing polymers which have no structural similarities to antifreeze proteins but reproduce the same macroscopic properties, potentially by different molecular-level mechanisms. The application of these polymers to the cryopreservation of donor cells is also introduced. Ice crystal growth is a major problem in cell and tissue cryopreservation for transplantation, transfusion, icing of aircraft wings and many other applications. Here the authors review the emerging field of synthetic macromolecular mimics of antifreeze proteins that can be used overcome such problems.

Ice formation and growth is of interest to many different fields, including food science, mechanical engineering, agriculture and cryobiology, however little is understood about the underlying mechanisms behind the nucleation and growth processes. The need to increase our understanding of ice and how it is affected by compounds with 'antifreeze' properties is fundamental to improving techniques for the storage of biologics. Nature has evolved to contend with a range of harsh climates; in particular, they produce cryoprotectants enabling them to survive sub-zero temperatures. Inspired by Nature's ingenious response a range of synthetic protein mimics have been developed, which have ice growth inhibition activity. The scientific principles behind ice nucleation and growth, and the materials that affect them, as well as current techniques for analysis are detailed in Chapter 1. This thesis reports on ice-activity for a range of compounds, studying their micro- and macroscopic effects on ice, as well as any potential cryoprotective capabilities, with the view to further fundamental understanding of ice growth inhibition and aid in the development of future potent cryoprotectants. A diverse range of methods including microscopy, X-ray Diffraction (XRD) and solid state nuclear magnetic resonance (SSNMR) were used to aid in characterisation and analysis by monitoring structural changes as well as the antifreeze macromolecule:ice interface. Chapter 2 investigates cryostorage of a range of biological materials using an organic solvent-free formulation consisting of an ice growth inhibiting polymer and a secondary bulking agent. Chapter 3 details X-ray diffraction (XRD) as a new method for studying ice growth continuously as a function of time, confirming its potential as a supplementary tool to study ice growth. Chapter 4 builds upon results from microscopy and XRD-based methods by using solid state nuclear magnetic resonance (SSNMR) to enable the study of molecular-level details experimentally. SSNMR provides further evidence for the 'turning on' of ice recrystallisation activity (IRI) for poly(vinyl alcohol) and ice-binding for a variety of compounds. Chapter 5 focuses on ice nucleation specifically. A range of previously untested materials that feature design motifs associated with nucleators reported on in the literature were examined for ice nucleation effectiveness and IRI activity, finding none to inhibit ice growth, and that structure alone is not enough to infer nucleation effectiveness.