Cold denaturation as a tool to measure protein stability

Abstract

Protein stability is an important issue for the interpretation of a wide variety of biological problems but its assessment is at times difficult. The most common parameter employed to describe protein stability is the temperature of melting, at which the populations of folded and unfolded species are identical. This parameter may yield ambiguous results. It would always be preferable to measure the whole stability curve. The calculation of this curve is greatly facilitated whenever it is possible to observe cold denaturation. Using Yfh1, one of the few proteins whose cold denaturation occurs at neutral pH and low ionic strength, we could measure the variation of its full stability curve under several environmental conditions. Here we show the advantages of gauging stability as a function of external variables using stability curves.

Distribution of Protein Content and Number of Aggregates in Monoclonal Antibody Formulation After Large-Scale Freezing

“…Also, chemical degradation reactions, such as deamidation, hydrolysis, or oxidation are slowed down (3). Disadvantages include possible stress for proteins through cold denaturation (6,7) or pH-shifts due to freeze-concentration (8,9), to name just two. It had also been shown that the ice-liquid interface may pose a problem for proteins (10–12), e.g.…”

Impact of Buffer, Protein Concentration and Sucrose Addition on the Aggregation and Particle Formation during Freezing and Thawing

“…Like all other therapeutic proteins, monoclonal antibodies (mAb) may undergo structural changes during FT processes which lead to protein aggregation, misfolding/unfolding, loss of biological activity or enhanced immune response in patients ( 5 – 7 ). The aggregation process is generally poorly understood ( 8 ) and may have several causes, such as cryoconcentration ( 9 ), contact with interfaces ( 1 , 10 , 11 ), cold denaturation ( 12 , 13 ), and many others ( 14 – 17 ).…”

Effect of temperature during vital gluten adhesive preparation and application on shear-bond strength

“…Recent studies suggest that the hydrophilic effect is much more important than the hydrophobic effect in cold denaturation, assuming that the hydrophilic interactions are the predominant forces for the folding of proteins and enabling protein solubility [11,12]. The stability of proteins is probably a dual concept involving both thermodynamic stability and thermal resistance, and denaturation occurs as thermal denaturation at high temperatures and cold denaturation at low temperatures [13].…”