Chapter 3. Effect of mechanical stresses on the native structure of proteins
3.6 Conclusion
In this chapter, the effects of mechanical stresses on protein native structure was studied. The strongest deleterious effects of agitation on the native structure of protein was observed in samples subjected to high shear from mixing in hypodermic needle. This was considered a simulated real conditions in industry during ampoule or vial filling and during mixing in prefilled syringes for use by patients. For long pipe filling systems, it is important to understand the force the relative effects when applied to the formulation (in liquid). Very high levels of pressure can create high shear stress and produce protein aggregates. Reducing the length of the pipe and filling speed as well as increasing the diameter of the pipe are advantageous in reducing potential for aggregation/denaturation.
Although the degree of aggregation in samples prepared by low shear agitation (magnetic stirring) is relatively low compared to the other agitation methods, it is essential for the manufacturer or protein user to know that the presence of any particles may act as a nuclei resulting in nucleation and growth mechanism of aggregation. This experimental setup can be a useful tool to monitor flow-induced protein aggregation with high reproducibility since all flow materials experience varied degrees of shear.
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