Work

Complete and Rapid NMR Characterization of Therapeutic Monoclonal Antibodies via Differential Labeling of Fab Fragments in Escherichia coli

Year: 2026

Type: article

Abstract: to produce the heavy and light chains separately, thereby allowing differential labeling of either chain of the refolded fragment. This was demonstrated on the Fab fragments of adalimumab, bevacizumab, infliximab, rituximab, and trastuzumab. The new labeling scheme produced NMR spectra with almost no resonance overlap, allowing near-complete assignments (>90%) of backbone resonances in less than an hour. In addition, a near-complete assignment of side-chain methyl groups was carried out that allowed the assignments of Avastin. The total time from Petri dish to final backbone assignment was 5 weeks per chain.

Source: Analytical Chemistry

DOI: 10.1021/acs.analchem.6c00109

Authors: Donald Gagné, Yves Aubin

Institutions: Health Canada, Carleton University

Language: English

Topic: Advanced NMR Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: hybrid

Estimated APC paid: $4,500

Funder: Government of Canada