Protein Stability: How To Analyze It Effectively?
Protein stability is one of the common problems in protein expression. This issue is a major problem in protein purification, storage, and formulation as well. Consequently, we are going to explain protein stability in brief so that everyone can understand why it is a hot topic for the Medicine industry.
Typically, if the proteins are folded appropriately, the will remain stable throughout the protein expression and purification. As a result, you can extract enough protein. In contrast to this situation, some proteins that is unstable throughout the protein purification process. In fact, some are so unstable that you will not be able to obtain any of them.
Several factors play a significant role in this volatile situation such as the amino acid sequence of the protein, host cell strain, protein construction. Due to these factors, the protein expression, and purification process becomes complicated.
Furthermore, the folded proteins are slightly stable under physiological conditions. Different other forces like electrostatic interactions, hydrophobic effects, and hydrogen bonding work as stabilizing factors. They are the primary driving forces for the protein folding process.
How To Analyze Protein Stability?
Several techniques have been used to determine whether a protein is stable or not. To make an analysis, different spectroscopic ways like – UV, fluorescence, and infrared are utilized. On the other hand, the essential thermodynamic processes, for example – differential scanning calorimetry (DSC) are used to measure the effect of temperature on protein stability.
Besides, the all-important peptide-mapping process is another significant tool to find out the chemical changes within a protein like deamidation or oxidation.
Aside from the above techniques, you can also use HPLC to analyze the purity of protein. Other than these, many experts use various methods like SDS-PAGE, capillary electrophoresis, and isoelectric focusing to measure the stability. On top of it, you can determine the state of aggregation with particle size and arrayed instruments.
To put it simply, you will find many processes of measuring the protein stability. Nevertheless, all these complex processes suggest that analyzing the stability of a protein is not an easy task. Above all, keeping nature of protein structure intact is by far the most important factor for achieving any success with biopharmaceutical product.