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Related Experiment Video

Updated: Nov 7, 2025

Author Spotlight: Experimental Approaches for the Synthesis of Low-Valent Metal-Organic Frameworks from Multitopic Phosphine Linkers
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Pragmatic mAb lead molecule engineering from a developability perspective.

Bertie Chi1, Geoffrey De Oliveira1, Tom Gallagher1,2

  • 1Kymab Ltd., Cambridge, UK.

Biotechnology and Bioengineering
|April 29, 2021
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Summary
This summary is machine-generated.

Antibody developability is crucial for drug success. This study identified and corrected specific mutations causing precipitation, improving manufacturability without affecting biological activity.

Keywords:
developabilityforced precipitationmAb purificationmutagenesissolubility

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Area of Science:

  • Biopharmaceutical development
  • Protein engineering
  • Molecular biology

Background:

  • Increasing number of approved antibody drugs highlights the importance of "developability" for successful drug candidates.
  • Developability encompasses both biological potency and manufacturability attributes, requiring simultaneous selection of molecules with both.
  • Antibody precipitation during purification is a significant challenge in bioprocess development.

Purpose of the Study:

  • To identify and engineer mutations to improve the developability profile of an antibody exhibiting problematic precipitation.
  • To investigate the hypothesis that unusual positively charged mutations at the VH-VL interface contribute to precipitation.
  • To assess the impact of reversions on expression, stability, and biological activity.

Main Methods:

  • Bioinformatic analysis to identify potential sequence or structural liabilities.
  • Analysis of antibody light chain mutations at the VH-VL domain interface.
  • Reversion of identified residues to their germline sequence.
  • Forced precipitation assay to evaluate stability.
  • Assessment of expression titers and biological activity.

Main Results:

  • Two unusual positively charged mutations on the light chain were identified as likely contributors to precipitation.
  • Reversion of these residues to germline significantly improved expression titers and stability in precipitation assays.
  • No loss of biological activity was observed in the reverted mutants.
  • Bespoke process optimization for large-scale manufacturing was also successful.

Conclusions:

  • Context-dependent sequence liabilities, such as the identified mutations, should be incorporated into early in silico developability screening.
  • Efficient mitigation of such liabilities is possible without extensive screening of lead molecule variants.
  • Improving antibody developability through targeted mutagenesis enhances manufacturability and streamlines drug development.