Multimeric VHH
Nanobody Expression
BACKGROUND
- VHH single domain antibodies readily express in the Pfenex Expression Technology®
- Expression and enrichment of a variety of VHH specificities facilitated early drug development
Challenge
Assess VHH concatemer expression and linker strategies to enhance function, avidity, and half-life in the Pfenex Expression Technology®
Solution
Hundreds of unique strains screened at 96-well scale for optimized expression of VHH 6X His-tagged concatemers
3X VHH Multimer Expression
- A unique 3X VHH concatemer expression strategy evaluated with multiple Pfenex toolbox secretion leaders
- Freeze-thaw lysis method developed for periplasmic release of concatemers
- 96-well plate IMAC purification developed to enrich His-tagged concatemers
- Best secretion leader/linker strategy enriched >2 mgs of 3X concatemer from one 96-well plate
Key Results
- Strains were identified expressing 3X, 4X, and 5X multimers utilizing secretion leaders specific for periplasmic localization of the VHH
- Cell-based assays using enriched multimeric VHH confirmed functional activity
