Higher order structure (HOS) is critical for product and process understanding in biopharmaceutical development. The clinical effect and biological functions of biologics are closely related to HOS which can be affected by processing, formulation and storage conditions. Therefore, HOS is important to determine structure function relationship and guide process development for biologics. HOS determination is challenging for large proteins like antibodies. Different biophysical tools have different levels of resolution and throughput. Light Scattering for protein size and Circular Dichroism for secondary structure provide low resolution structural information. X-ray Crystallography and Nuclear Magnetic Resonance have high resolution but are not suitable for large proteins. Mass Spectrometry and Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) are increasingly used to determine protein sequence and conformation but need special expertise and have low throughput. Protein Conformational Array (PCA) is a new technique to determine protein HOS. This ELISA or Luminex<sup>®</sup>-based method uses a panel of 34 antibodies to detect epitopes that cover the whole antibody and thus indicate the antibody 3-D structure. It is simple to perform with high throughput and can be a complementary technique to HDX-MS. It was initially used to determine biosimilarity for antibodies but is being further developed as a process development tool for novel antibodies. In this study, PCA is used to predict the structures of antibody aggregates, compare antibodies expressed from difference cell lines, and monitor antibody structure changes caused by processing and different conditions. Protein Conformational Array is a very promising tool for antibody HOS determination to provide in-depth product and process understanding.<br/>

Protein conformational array as a process development tool for antibody higher order structure analysis

251st National Meeting (2016)

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