A dual-purpose protein ligand for effective therapy and sensitive diagnosis of anthrax.
This article reports the design of a bivalent protein ligand with dual use in therapy and diagnosis of anthrax caused by Bacillus anthracis. The ligand specifically binds to PA and thereby blocks the intracellular delivery of LF and EF toxins that, respectively, cause cell lysis and edema. The ligand is a chimeric scaffold with two PA-binding domains (called VWA) linked to an IgG-Fc frame. Molecular modeling and binding measurements reveal that the VWA-Fc dimer binds to PA with high affinity (K(D)=0.2 nM). An in vitro bio-luminescence assay shows that VWA-Fc (at nanomolar concentration) protects mouse macrophages from lysis by PA/LF. In vivo studies demonstrate that VWA-Fc at low doses (approximately 50 microg/animal) are able to rescue animals from lethal doses of PA/LF and B. anthracis spores. Finally, VWA-Fc is utilized as the capture molecule in the sensitive (down to 30 picomolar) detection of PA using surface plasmon resonance.
Vuyisich
Momchilo
M
Biosciences Division, Group B-7, Los Alamos National Laboratory, Los Alamos, NM 87545, USA.
Gnanakaran
S
S
Lovchik
Julie A
JA
Lyons
C Rick
CR
Gupta
Goutam
G
D000818:Animals; D000881:Anthrax / Q000175:diagnosis* / Q000628:therapy*; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D012680:Sensitivity and Specificity
D000942:Antigens, Bacterial; D008024:Ligands