Camelid single domain antibodies (VHHs) as neuronal cell intrabody binding agents and inhibitors of Clostridium botulinum neurotoxin (BoNT) proteases

Jacqueline M. Tremblay, Chueh Ling Kuo, Claudia Abeijon, Jorge Sepulveda, George Oyler, Xuebo Hu, Moonsoo M. Jin, Charles B. Shoemaker

Research output: Contribution to journalArticlepeer-review

72 Scopus citations

Abstract

Botulinum neurotoxins (BoNTs) function by delivering a protease to neuronal cells that cleave SNARE proteins and inactivate neurotransmitter exocytosis. Small (14 kDa) binding domains specific for the protease of BoNT serotypes A or B were selected from libraries of heavy chain only antibody domains (VHHs or nanobodies) cloned from immunized alpacas. Several VHHs bind the BoNT proteases with high affinity (KD near 1 nM) and include potent inhibitors of BoNT/A protease activity (Ki near 1 nM). The VHHs retain their binding specificity and inhibitory functions when expressed within mammalian neuronal cells as intrabodies. A VHH inhibitor of BoNT/A protease was able to protect neuronal cell SNAP25 protein from cleavage following intoxication with BoNT/A holotoxin. These results demonstrate that VHH domains have potential as components of therapeutic agents for reversal of botulism intoxication.

Original languageEnglish (US)
Pages (from-to)990-998
Number of pages9
JournalToxicon
Volume56
Issue number6
DOIs
StatePublished - Nov 2010

Keywords

  • BoNT
  • Botulinum
  • Intrabody
  • Metalloproteinase
  • Nanobody
  • Neurotoxin
  • VHH

ASJC Scopus subject areas

  • Toxicology

Fingerprint

Dive into the research topics of 'Camelid single domain antibodies (VHHs) as neuronal cell intrabody binding agents and inhibitors of Clostridium botulinum neurotoxin (BoNT) proteases'. Together they form a unique fingerprint.

Cite this