期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 53, 期 1, 页码 132-135出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201307939
关键词
antibodies; CDR; coiled coil; polypeptide; protein engineering
资金
- National Institutes of Health (NIH) [R01 GM097206]
The X-ray crystal structure of a bovine antibody (BLV1H12) revealed a unique structure in its ultralong heavy chain complementarity determining region3 (CDR3H) that folds into a solvent-exposed -strand stalk fused to a disulfide crosslinked knob domain. We have substituted an antiparallel heterodimeric coiled-coil motif for the -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.
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