This work was supported by the National Institutes of Health (NIH) Grant R01 GM097206 (to P.G.S.). This manuscript is number 25001 of The Scripps Research Institute.
An Antibody with a Variable-Region Coiled-Coil “Knob” Domain†
Article first published online: 19 NOV 2013
Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Angewandte Chemie International Edition
Volume 53, Issue 1, pages 132–135, January 3, 2014
How to Cite
Zhang, Y., Goswami, D., Wang, D., Wang, T.-S. A., Sen, S., Magliery, T. J., Griffin, P. R., Wang, F. and Schultz, P. G. (2014), An Antibody with a Variable-Region Coiled-Coil “Knob” Domain. Angew. Chem. Int. Ed., 53: 132–135. doi: 10.1002/anie.201307939
- Issue published online: 23 DEC 2013
- Article first published online: 19 NOV 2013
- Manuscript Received: 10 SEP 2013
- National Institutes of Health (NIH). Grant Number: R01 GM097206
- The Scripps Research Institute
- coiled coil;
- protein engineering
The X-ray crystal structure of a bovine antibody (BLV1H12) revealed a unique structure in its ultralong heavy chain complementarity determining region 3 (CDR3H) that folds into a solvent-exposed β-strand “stalk” fused to a disulfide crosslinked “knob” domain. We have substituted an antiparallel heterodimeric coiled-coil motif for the β-strand stalk in this antibody. The resulting antibody (Ab-coil) expresses in mammalian cells and has a stability similar to that of the parent bovine antibody. MS analysis of H–D exchange supports the coiled-coil structure of the substituted peptides. Substitution of the knob-domain of Ab-coil with bovine granulocyte colony-stimulating factor (bGCSF) results in a stably expressed chimeric antibody, which proliferates mouse NFS-60 cells with a potency comparable to that of bGCSF. This work demonstrates the utility of this novel coiled-coil CDR3 motif as a means for generating stable, potent antibody fusion proteins with useful pharmacological properties.