Optimized bacterial expression of a synthetic BRIL antibody

Journal article

The use of monoclonal fragments antigen binding (Fabs) is a prevalent methodology facilitating protein structure determination via both crystallography and cryo‐EM. The development of a synthetic Fab against the BRIL domain improved the accessibility of this approach, providing a general fiducial applicable to any protein of interest via the simple curation of a BRIL fusion protein. Here, we document the generation of a T7 Express ΔcybC strain allowing contaminant‐free bacterial expression of the synthetic anti‐BRIL Fab BAG2. We also report the crystal structure of BAG2 in complex with native cytochrome b562, a complex arising from expression in canonical Escherichia coli strains.

Authors: Cooper, BF; Isom, GL

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: International Union of Crystallography
• Journal: Acta Crystallographica Section F: Structural Biology Communications
• Volume: 82
• Issue: 4
• Publication date: 2026-03-17
• Acceptance date: 2026-02-13
• DOI: 10.1107/s2053230x26001548
• EISSN: 2053230X
• ISSN: 2053230X
• Language: English
• Keywords: cryo‐EM; synthetic antibodies; BRIL; fiducials; X‐ray crystallography