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The Discovery, Engineering and Characterisation of a Highly Potent Anti-Human IL-13 Fab Fragment Designed for Administration by Inhalation

Author:
Lightwood, Daniel, O'Dowd, Victoria, Carrington, Bruce, Veverka, Vaclav, Carr, Mark D., Tservistas, Markus, Henry, Alistair J., Smith, Bryan, Tyson, Kerry, Lamour, Sabrina, Sarkar, Kaushik, Turner, Alison, Lawson, Alastair D., Bourne, Tim, Gozzard, Neil, Palframan, Roger
Source:
Journal of Molecular Biology 2013 v.425 pp. 577-593
ISSN:
0022-2836
Subject:
atomization, binding capacity, breathing, epitope mapping, genes, humans, immunoglobulin G, in vitro studies, interleukin-13, intranasal administration, neutralization, nuclear magnetic resonance spectroscopy
Abstract:
We describe the discovery, engineering and characterisation of a highly potent anti-human interleukin (IL)-13 Fab fragment designed for administration by inhalation. The lead candidate molecule was generated via a novel antibody discovery process, and the selected IgG variable region genes were successfully humanised and reformatted as a human IgG γ1 Fab fragment. Evaluation of the biophysical properties of a selection of humanised Fab fragments in a number of assays allowed us to select the molecule with the optimal stability profile. The resulting lead candidate, CA652.g2 Fab, was shown to have comparable activity to the parental IgG molecule in a range of in vitro assays and was highly stable. Following nebulisation using a mesh nebuliser, CA652.g2 Fab retained full binding affinity, functional neutralisation potency and structural integrity. Epitope mapping using solution nuclear magnetic resonance confirmed that the antibody bound to the region of human IL-13 implicated in the interaction with IL-13Rα1 and IL-13Rα2. The work described here resulted in the discovery and design of CA652.g2 human γ1 Fab, a highly stable and potent anti-IL-13 molecule suitable for delivery via inhalation.
Agid:
1021732