Ice crystallization causes cellular and material damage in subzero environments, presenting challenges in medicine, food preservation, and cryopreservation. Antifreeze proteins (AFPs) are specialized polypeptides produced by cold-adapted organisms that inhibit ice growth by binding specific ice planes and inducing thermal hysteresis without altering melting points. This project aims to purify and characterize AFPs from Zoarces americanus, Lycodichthys dearborni, Pseudopleuronectes americanus, and Daucus carota recombinantly expressed in Escherichia coli. We will express and purify AFP proteins using immobilized metal ion affinity chromatography. We plan to evaluate the function of these AFP proteins in cryopreservation, food preservation, and anti-icing applications. We have developed preliminary assays including freeze–thaw survival testing in E.coli and injections in Eisenia fetida to assess tolerance for future cryopreservation studies. We utilized initial thermal hysteresis assays with glycerol as a cryoprotectant, which demonstrated reduced freezing rates. Our next steps will involve evaluating purified AFP proteins with our various frameworks targeted to control ice formation in biological and industrial systems.