Development and scale-up of the recovery and purification of a domain antibody Fc fusion protein-comparison of a two and three-step approach

Biotechnol Bioeng. 2015 Jul;112(7):1417-28. doi: 10.1002/bit.25561. Epub 2015 Mar 13.

Abstract

A robust, economical process should leverage proven technology, yet be flexible enough to adopt emerging technologies which show significant benefit. Antibody and Fc-fusion processes may capitalize on the high selectivity of an affinity capture step by reducing the total number of chromatographic steps to 2. Risk associated with this approach stems from the potentially increased time frame needed for process development as well as unforeseen changes in impurity profile during first scale-up of drug substance (DS) for animal toxicology and clinical phase I trials (FIH) production, which could challenge a two-step process to the point of failure. Two different purification strategies were pursued during process development for an FIH process of a dAB-Fc fusion protein. A two-step process was compared to a three-step process. The two-step process leveraged additives to maximize impurity reduction during affinity capture. While wash additives in combination with a mixed mode chromatography met all impurity reduction requirements, HCP levels were still higher as compared to the three-step process. The three-step process was implemented for manufacture of clinical material to mitigate risk.

Keywords: Fc-fusion protein; caprylate; design of experiments; process development; purification; scale-up.

MeSH terms

  • Animals
  • CHO Cells
  • Chromatography, Liquid / methods*
  • Cricetulus
  • Immunoglobulin Fc Fragments / genetics
  • Immunoglobulin Fc Fragments / isolation & purification*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification*

Substances

  • Immunoglobulin Fc Fragments
  • Recombinant Fusion Proteins