Get access

An Antibody with a Variable-Region Coiled-Coil “Knob” Domain

Authors

  • Dr. Yong Zhang,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    2. Present address: California Institute for Biomedical Research (Calibr), 11119 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    Search for more papers by this author
  • Dr. Devrishi Goswami,

    1. Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, FL 33458 (USA)
    Search for more papers by this author
  • Dr. Danling Wang,

    1. California Institute for Biomedical Research (Calibr), 11119 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    Search for more papers by this author
  • Dr. Tsung-Shing Andrew Wang,

    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    Search for more papers by this author
  • Shiladitya Sen,

    1. Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210 (USA)
    Search for more papers by this author
  • Dr. Thomas J. Magliery,

    1. Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210 (USA)
    Search for more papers by this author
  • Dr. Patrick R. Griffin,

    1. Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, FL 33458 (USA)
    Search for more papers by this author
  • Dr. Feng Wang,

    Corresponding author
    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    2. Present address: California Institute for Biomedical Research (Calibr), 11119 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    • Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)

    Search for more papers by this author
  • Dr. Peter G. Schultz

    Corresponding author
    1. Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    2. Present address: California Institute for Biomedical Research (Calibr), 11119 N. Torrey Pines Road, La Jolla, CA 92037 (USA)
    • Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (USA)

    Search for more papers by this author

  • 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.

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.

Get access to the full text of this article

Ancillary