Pfast™ Lead Identification and Optimization Supporting Computational Drug Discovery Technology 

by | Aug 24, 2026 | Case Studies

Pfast™ Lead Identification and Optimization

Supporting Computational Drug Discovery Technology

Pfenex Expression Technology® | Primrose Bio

Challenge

  • AI generated, humanized Fab from partner required production and binding assessment
  • A platform approach was requested by partner for evaluating this and future programs using this new workflow

Expression Strategy

  • Design coding sequences for diverse Fab designs to identify orientation, secretion leaders, and RBS combinations to work for three similar Fab variants
  • Workflow developed to optimize expression and purification of produced proteins in an automated 96-well scale format
  • Soluble lysate fractions analyzed by SDS-CGE (non-reduced) to verify Fab assembly

Results & Takeaways

  • Soluble expression >1,000mcg/mL for multiple Fab variants achieved.
  • 24 high-expressing Fabv1 strains (>350mcg/mL soluble titer) enable identification of Fab expression strategies.
  • Selected expression strategies used to screen Fab variants with soluble expression ranging to >1,000mcg/mL.
  • Antigen binding of 48 Fab constructs confirmed by ELISA.
  • This simplified method accelerates drug discovery pipelines’ generation of humanized Fab molecules with improved antigen binding.
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