Structural Characterization of Recombinant Therapeutic Proteins by Circular Dichroism

Carlo      Bertucci; Marco      Pistolozzi; Angela      De Simone

doi:10.2174/138920111798357276

Abstract

Most of the protein therapeutics are now produced by recombinant DNA technology. The advantages of recombinant proteins are related to their higher specificity and to their safety as exposure to animal or human diseases. However, several problems are still present in development of recombinant proteins as therapeutics, such as low bioavailability, short serum half-life, and immune response. Their successful application hinges on the protein stereochemical stability, and on the folding and the tendency to aggregate induced by purification steps and storage. All these aspects determine the failure of many potential protein therapies, and limitations in the development of the formulation. The application of multiple analytical techniques is important in order to obtain a detailed product profile and to understand how manufacturing can influence product structure and activity. Surely the protein conformation is a key aspect to be assessed, because a specific conformation is often essential for the biological function of the protein. Thus, there is a growing need to perform structural studies under the conditions in which the proteins operate, and to monitor the structural changes of the protein. Circular dichroism has been increasingly recognised as a valuable and reliable technique to get this information. In particular, examples will be here reported on the use of circular dichroism spectroscopy in the structural characterization of free and formulated recombinant proteins, looking at the prediction of the secondary structure, propensity to conformational changes, stability, and tendency to aggregate.

Keywords: Aggregation, bioavailability, circular dichroism, protein formulation, quality control, recombinant protein therapeutics, thermal stability, recombinant DNA technology, short serum half-life, protein stereochemical stability, circular dichroism spectroscopy in, conformational changes, tendency to aggregate, gene therapy, efficacy