Purification of ADCs by Hydrophobic Interaction Chromatography

  • Calvin L. BeckerEmail author
  • Robert J. Duffy
  • Jorge Gandarilla
  • Steven M. Richter
Part of the Methods in Molecular Biology book series (MIMB, volume 2078)


Antibody-drug conjugate (ADC) in vitro potency has been shown to be dependent on drug load, with higher drug load providing lower IC50 values. However, in vivo potency is affected by intrinsic biological effects as well, such as plasma clearance, dose-limiting toxicity, etc. Developing a preparative HIC process for ADC purification to isolate species with a specific drug loading involves several steps including conjugation optimization, resin selection, solubility studies gradient screening, and step gradient development (buffer selection). In this chapter, the rationale and general considerations for developing a preparative hydrophobic interaction chromatography (HIC) method are described for isolation of an example ADC with specific drug load, e.g., two monomethyl auristatin E (MMAE) payloads (E2).

Key words

Antibody-drug conjugate (ADC) Hydrophobic interaction chromatography (HIC) Drug-antibody ratio (DAR) Lyotrope Buffers Gradient 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Calvin L. Becker
    • 1
    Email author
  • Robert J. Duffy
    • 2
  • Jorge Gandarilla
    • 1
  • Steven M. Richter
    • 1
  1. 1.Abbvie Inc.North ChicagoUSA
  2. 2.Abbvie Inc.Redwood CityUSA

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