Ultrafast Separation and Analysis of Monoclonal Antibody Aggregates Using Membrane Chromatography

Anal Chem. 2017 Apr 18;89(8):4716-4720. doi: 10.1021/acs.analchem.7b00580. Epub 2017 Apr 6.

Abstract

We discuss a method for rapid and cost-effective analysis of monoclonal antibody (mAb) aggregates. Hydrophobic interaction membrane chromatography, which was previously shown to be highly suitable for such separation and analysis, was used in a recently developed format referred to as laterally fed membrane chromatography (or LFMC). A stack of rectangular polyvinylidene fluoride (or PVDF) membranes having 0.22 μm pores housed within a modified analytical-scale LFMC device was used for analyzing aggregate types and content in different monoclonal antibody samples. High-resolution separations could be achieved in less than 1.5 min, this being faster than other currently available techniques such as size exclusion ultraperformance liquid chromatography (SE-UPLC). Moreover, the operating pressure was less than 200 kPa, which eliminated the need for an expensive high-pressure pump and chromatography system. The resolution obtained using the LFMC was comparable to that obtained using SE-UPLC. The effect of design variations such as change in dead volume and pillar size within the lateral channels within the LFMC device was also examined.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / metabolism
  • CHO Cells
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid*
  • Cricetinae
  • Cricetulus
  • Dimerization
  • Hydrophobic and Hydrophilic Interactions
  • Polyvinyls / chemistry
  • Protein Aggregates*

Substances

  • Antibodies, Monoclonal
  • Polyvinyls
  • Protein Aggregates
  • polyvinylidene fluoride