Hydrophobic Interaction Process Scale Separation of Protein, MAb and Antibody Drug Conjugate

Haiying Chen, R&D, Sepax Technologies, Inc., Delaware, USA

Hydrophobic interaction chromatographic (HIC) separation applies to all stages of the biological sample purification process, include capturing and polishing. Different forms of proteins, aggregates, and fragments can be isolated due to the hydrophobic interaction between the sample and the resin solid support. This hydrophobic interaction can be controlled by a number of factors such as protein structure, salt concentration, pH, temperature and organic solvent additives. HIC separation is a great addition to the native protein purification process including ion exchange, size exclusion, and affinity chromatography. Generik & Polar MC-HIC resins are introduced for protein, mAb and antibody drug conjugate process scale purification. The polymeric resins are composed of a methacrylate polymer bead (also referred to as PMA) with high mechanical and chemical stability. The resins have mean particle size of 30 and 60 µm respectively, and pore size of 800 Å. On the resin surface, alkyl groups or aryl groups are attached via proprietary chemistry, so the resin selectively interacts with bio-molecules by utilizing the variations in hydrophobicity. The characteristics for the 30 µm particle size are discussed for the Generik & Polar MC-HIC resins. Protein separation examples are presented for the resin applications. Sepax Generik MC30-HIC Butyl, Polar MC30-HIC Butyl and Polar MC30-HIC Ether resins provide excellent high efficiency and recovery separation of bio-molecules. Monoclonal antibody, ADC (antibody drug conjugate), DNA, and other oligonucleotides are all separated via the hydrophobic interaction. The HIC media also tolerates high-pressure operation up to 100 bars. All three phases are applicable at different stages from laboratory discovery, pilot-scale purification to industrial process chromatography.