Multimodal Chromatography for Purification of Biotherapeutics - A Review

Curr Protein Pept Sci. 2019;20(1):4-13. doi: 10.2174/1389203718666171020103559.


Process chromatography forms the core of purification of biotherapeutics. The unparalleled selectivity that it offers over other alternatives combined with the considerable robustness and scalability make it the unit operation of choice in downstream processing. It is typical to have three to five chromatography steps in a purification process for a biotherapeutic. Generally, these steps offer different modes of separation such as ion-exchange, reversed phase, size exclusion, and hydrophobic interaction. In the past decade, multimodal chromatography has emerged as an alternative to the traditional modes. It involves use of more than one mode of separation and typically combines ion-exchange and hydrophobic interactions to achieve selectivity and sensitivity. Over the last decade, numerous authors have demonstrated the significant potential that multimode chromatography offers as a protein purification tool. This review aims to present key recent developments that have occurred on this topic together with a perspective on future applications of multimodal chromatography.

Keywords: Biopharmaceuticals; Fab; MMC; Multimodal chromatography/mixed-mode chromatography; antibody fragments; host cell proteins; mAb; process related impurities; product related impurities; therapeutic proteins..

Publication types

  • Review

MeSH terms

  • Animals
  • Antibodies, Monoclonal / isolation & purification*
  • Bacteria / chemistry
  • Biological Products / isolation & purification*
  • Chromatography / instrumentation
  • Chromatography / methods*
  • Humans
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions
  • Immunoglobulin Fab Fragments / isolation & purification*
  • Protein Binding
  • Proteins / isolation & purification*
  • Resins, Synthetic / chemistry*
  • Solvents / chemistry
  • Static Electricity


  • Antibodies, Monoclonal
  • Biological Products
  • Immunoglobulin Fab Fragments
  • Proteins
  • Resins, Synthetic
  • Solvents