Biophysical Techniques For Characterizing The Higher Order Structure And Interactions Of Monoclonal Antibodies

One of the major challenges that the biopharmaceutical industry currently faces is maintaining the structural integrity and stability of protein therapeutics through production and shelf life. Changes introduced in antibody production processes or formulations may have major impact on stability, efficacy, and safety. Hence, a robust set of biophysical techniques to characterize and assess the "higher order structure" (HOS) of monoclonal antibodies (mAbs) is an integral part of the drug development process. In a unique collaboration spanning academic/government institutions and the biotechnology industry, we used a set of sophisticated orthogonal biophysical techniques for characterizing the HOS of an IgG1 mAb. Each section of this chapter focuses on a particular method to probe different molecular properties of the mAb such as solution mass, size, polydispersity, solution charge, thermal stability, aggregation, and secondary and tertiary structural changes in different solutions and at different mAb concentrations. Our collaboration has resulted in the collection of biophysical data rich in the solution structural information of the NISTmAb that we hope will help researchers worldwide as a guide for the biophysical characterization of mAbs and proteins in general.