Prevent Protein Aggregation in Biopharma Formulations

The structure of biomolecules, such as monoclonal antibodies and other proteins or peptides, is highly relevant for their therapeutic effect. Under certain conditions, biomolecules are highly susceptible to aggregation and unfolding which leads to reduced stability and a loss of functionality. Elevated temperatures, mechanical stress during manufacturing and transport, extreme pH conditions, and high protein concentrations can lead to partial unfolding of proteins, resulting in self-association and aggregates that compromise therapeutic activity and patient safety.

To enhance biomolecule stability, it is essential to ensure optimal formulation and environmental conditions such as optimal pH, avoiding high and low temperatures, and minimizing shear stress. Using stabilizing excipients like sugars, polyols, and amino acids can protect against thermal and mechanical stress, reducing the risk of protein aggregation during manufacturing, storage and administration.

Sugars and sugar alcohols (e.g., sucrose, trehalose, myo-inositol, mannitol) are commonly used as stabilizers in liquid formulations or during lyophilization. During lyophilization, they form a glass matrix that replaces water in hydrogen bonds, maintaining the protein's native structure. In solution, they shift the equilibrium towards the protein’s native folded state by strengthening the hydration shell or binding to the unfolded protein. Read our case study.

Proteins can form a film at hydrophobic interfaces, which is susceptible to rupture (e.g. through mechanical stress like shaking of the vial) and subsequent aggregation. Surfactants, such as polysorbates and poloxamers, stabilize proteins by preventing adsorption to these surfaces and binding to hydrophobic sites of the protein molecule. This keeps the protein in solution and lessens its susceptibility to mechanical stress. Learn more in our case study.

Amino acids can serve multiple roles in biopharmaceutical formulations, acting as stabilizers, buffering agents, and antioxidants. They help maintain the protein's native structure and can solubilize the biomolecule. Amino acids known for their stabilizing effects include glycine, arginine, histidine and amino acid derivative N-acetyl-DL-tryptophan.