What is protein lyophilization?

Lyophilization involves freezing a protein solution followed by removal of ice by sublimation. The process subjects the protein to stresses such as denaturation at the ice surface, pH shifts and freeze concentration (Anchordoquy & Carpenter, 1996; Bhatnagar, Bogner, & Pikal, 2007; Chang, Kendrick, & Carpenter, 1996b).

What is meant by lyophilization of proteins and why is it done?

Lyophilization, or freeze-drying, is a method for the preservation of labile materials in a dehydrated form. It can be particularly suitable for high-value biomolecules such as proteins.

How do you calculate protein aggregation?

Biochemical assays for monitoring protein aggregates often rely on ultracentrifugation, size-exclusion chromatography, gel electrophoresis, dynamic light scattering, or turbidity measurements.

Why should proteins be lyophilized?

The fundamental goal of lyophilization is to impart desirable characteristics into the product, such as long-term stability, short reconstitution time, maintenance of the characteristics of the original liquid formulation upon reconstitution (e.g., solution properties, isotonicity, structures or conformation of …

What is lyophilization process?

Lyophilization or freeze drying is a process in which water is removed from a product after it is frozen and placed under a vacuum, allowing the ice to change directly from solid to vapor without passing through a liquid phase.

How do you identify aggregates?

For example, aggregates may be detected by orthogonal microscopy, chromatography or centrifugation methods. Undissolved species (other than gas bubbles or droplets) that are unintentionally present in the product. Particles can be foreign (not intrinsic to drug substance) or protein-related (i.e. large aggregates).

What is lyophilization technique?

Where is lyophilization used?

Lyophilization is also used in the biotechnology and biomedical industries to preserve vaccines, blood samples, purified proteins, and other biological material. This short laboratory procedure can be used with any commercially available freeze dryer to preserve your culture collection.

Does protein structure matter for lyophilization-induced aggregation?

Lyophilization can induce aggregation in therapeutic proteins, but the relative importance of protein structure, formulation and processing conditions are poorly understood. To evaluate the contribution of protein structure to lyophilization-induced aggregation, fifteen proteins were co-lyophilized with each of five excipients.

How important are protein descriptors in lyophilization of therapeutic proteins?

Results show that descriptors used for solution can also be useful in lyophilized systems. Lyophilization can induce aggregation in therapeutic proteins, but the relative importance of protein structure, formulation and processing conditions are poorly understood.

Can heuristics predict the aggregation of proteins in the lyophilized state?

The correlations show that descriptors obtained from solution-based heuristic methods can be used to quantitatively predict aggregation of proteins in the lyophilized state.

What determines the percent monomer remaining following lyophilization?

Within a given type of formulation, the percent monomer remaining following lyophilization is highly correlated to descriptors of protein structure pooled from the AGGRESCAN and PASTA algorithms along with a limited list of physical descriptors.