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An Antibody with a Variable‐Region Coiled‐Coil “Knob” Domain
Author(s) -
Zhang Yong,
Goswami Devrishi,
Wang Danling,
Wang TsungShing Andrew,
Sen Shiladitya,
Magliery Thomas J.,
Griffin Patrick R.,
Wang Feng,
Schultz Peter G.
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201307939
Subject(s) - antiparallel (mathematics) , coiled coil , antibody , complementarity determining region , fusion protein , structural motif , chemistry , microbiology and biotechnology , biophysics , recombinant dna , biology , immunoglobulin light chain , biochemistry , genetics , gene , physics , quantum mechanics , magnetic field
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.