Development of robust and highly efficient BIO-antifreezes for long-term biopreservation

The National Institute of General Medical Sciences awarded CAL State LA University Auxiliary Services Inc. $150,000 on August 1, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to develop robust and highly efficient bio-antifreezes for long-term biopreservation. The project runs through March 31, 2030, with performance in Los Angeles, California.

The research systematically determines the stability of key components within bio-antifreeze systems—specifically insect antifreeze proteins paired with small molecule enhancers—and pairs those analyses with quantitative assessment of functional activity. The work addresses a critical gap in understanding how antifreeze protein stability regulates enhancing activity in bio-antifreeze systems and their overall effectiveness. Success will provide mechanistic insights to guide rational development of cryoprotectant agents less toxic and more effective than current alternatives for long-term storage of cells and biological materials.

Abstract

The identification of less toxic and more effective cryoprotectant agents (cpas) is essential to enable the long- term storage of cells and other biological materials for human health. Produced in many cold-adapted organisms, antifreeze proteins (afps) have been recognized as natural antifreezes and cryoprotectants accounting for these organisms' survival in a subzero temperature environment. Afps exhibit great structural diversity, while are all characterized by their unique capabilities of non-colligative freezing-point depression and ICE recrystallization inhibition (IRI). The difference between the melting and depressed freezing points of water in the presence of afps, referred to as thermal hysteresis (TH), is generally used as a measure of the antifreeze activity of afps. Insect afps can exhibit 10-100 times greater TH activity particularly in the presence of enhancers than fish afps and small molecules are the more practical choice due to their greater stability, lower cost, and natural abundance at high concentrations compared to proteins. Advanced have been made in identifying efficient BIO-antifreeze systems (insect AFP and small molecule enhancer). Although many identified enhancers are known protein stabilizers, how the AFP's stability of regulates its enhancing activity in the BIO-antifreeze system and the overall effectiveness of the systems remains a critical GAP. The proposed research will systematically determine the key component's stability across the well-defined BIO-antifreezes and pairing these analyses with the quantitative assessment of functional activity of the systems. The completion of the project will reveal the mechanistic relationship between the two key characteristics of the BIO- antifreeze systems. Its success will provide foundational insights to guide the rational development of robust and highly efficient BIO-antifreezes for long-term biopreservation. Undergraduate and master's students will be fully integrated into this research under structured supervision. This sure project will thus significantly enhance and expand the research capacity at CAL state LA, a resource-limited institution, toward its sustainable research and education excellence.