An antibody with a variable-region coiled-coil "knob" domain
Journal
Angewandte Chemie - International Edition
Journal Volume
53
Journal Issue
1
Pages
132-135
Date Issued
2014
Author(s)
Zhang, Y.
Goswami, D.
Wang, D.
Sen, S.
Magliery, T.J.
Griffin, P.R.
Wang, F.
Schultz, P.G.
Abstract
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.
Type
journal article