ISBN 9783843956147

978-3-8439-5614-7, Reihe Mikrosystemtechnik

Hye In Kim
Prevention of heat and moisture induced protein denaturation through polymer intervention

192 Seiten, Dissertation Albert-Ludwigs-Universität Freiburg im Breisgau (2024), Softcover, B5

Zusammenfassung / Abstract

This study explores the use of neutrally charged, water-soluble acrylamide-based polymers as protein stabilizers under harsh conditions, such as elevated temperatures (≥ 60 °C), high humidity, and in solution. Designed to replace traditional stabilizers such as sugars, proteins, amino acids, and polyols, these polymers offer tunability and broader applicability across diverse proteins. Further, the role of hydrogen bonding was investigated with the polymers to better understand how amorphous additives preserve enzyme activity under stress.

Polymers with varying monomer ratios were characterized for glass transition temperature (Tg) and hygroscopicity to establish a customizable stabilization system. All polymers exhibited Tg values well above typical storage temperatures, with notable differences in hygroscopicity, suggesting variable interactions with enzymes. These polymers outperformed commonly used bovine serum albumin (BSA) in stabilizing reverse transcriptase and T4 DNA ligase under heat and humidity. Overall, acrylamide-based polymers show strong potential as versatile and effective protein stabilizers.