Decoding Protein Durability: Novel Stability and Storage Strategies

Abstract

Proteins are crucial molecules of biological processes with particular functions. They are marginally stable biomolecules and are readily prone towards denaturation due to the occurrence of different stressed conditions in solution, dried, or frozen forms. Some factors, such as pH, storage containers, temperature, buffer systems, and repetitive freezing and thawing, play a critical role in maintaining the stability of proteins. This review summarizes the pragmatic methods for the storage of purified proteins, preserving an ample amount of biological activity. It elucidates the simple and empirical means of preventing proteolytic denaturation, microbial contamination, low-temperature storage, and the inclusion of several kinds of stabilizing agents. Various types of protease inhibitors are known to reduce proteolytic degradation with improved protein storage stability. The addition of antimicrobials (e.g., thiomersal and sodium azide) and filtration are used to prevent microbial contamination. Low temperature and freeze-drying (lyophilization) are also known as significant methods for protein storage. Additionally, the inclusion of stabilizing agents, such as osmolytes, reducing agents, ionic stabilizers, specific substrates, ligands, and exogenous proteins, and concentrating diluted proteins are effective strategies for reducing the degradation and enhancing the shelf-life of proteins. Several stabilizing agents, such as -mercaptoethanol, EDTA, EGTA, glycerol, and ethylene glycol, enhance protein stability with less to no damage. Thus, these strategies provide substantial directions for maintaining the integrity of proteins, facilitating both investigational reproducibility and commercial viability of protein-based products.