Cat. No.
MABL-537
Application
neutralize, FC
Isotype
Engineer antibody
Species Reactivity
Clostridium
Clone No.
CR1
From
Recombinant Antibody
Specificity
CR1 binds to both BoNT/A1 and BoNT/A2 with high affinity (CR1 scfv: KD BoNT/A1 ~ 0.13 nM; KD BoNT/A2 ~ 0.4 nM. CR1 IgG: KD BoNT/A1 ~ 2.5 pM; KD BoNT/A2 ~ 1.7 nM.). CR1 Fab fragment binds to the BoNT/A1 binding domain, at the interface between the N-terminal lectin subdomain (HCN) and the C- terminal trefoil subdomain (HCC) at a discontinuous epitope consisting of β23-β24, β25-β26, β27-β28 and β35-β36 loops of the lectin subdomain and the C-terminus of the trefoil. Botulinum neurotoxin (BoNT) is secreted by Clostridial bacteria, and causes botulism which is characterized by flaccid paralysis and can be fatal without hospitalization. BoNT acts by inhibitnig acetylcholine release through binding with high affinity to SV2 on the presynaptic membrane and inducing its internalization by receptor-mediated endocytosis.
Alternative Names
BoNT/A; BoNT; Botulinum neurotoxin; Bontoxilysin-A; Bontoxilysin A; BOTOX
UniProt
P10845
Immunogen
CR1 was produced by a yeast gene diversity library used to increase the affinitiy of scFv AR2 for BoNT/A2, while maintinaing high affinity binding to BoNT/A2. AR2 was generated after multiple rounds of affinity maturation of the humanized C25, which itself was created by CDR-grafting from the parental murine C25. Murine C25 was prepared by constructing a scFv phage display library from mice immunized with recombinant BoNT/A binding domain (Hc).
Application Notes
Antibody binding can be studied by FC and the antibody also shows in vivo neutralizing activity against botulinum neurotoxin - the neutralizing activity is increased when administered in combination with the human BoNT/A antibodies RAZ1 and 2G11.
Antibody First Published
Garcia-Rodriguez et al. Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin. Nat Biotechnol. 2007 Jan;25(1):107-16.PMID17173035
Note on publication
Describes the process of increasing AR2 antibody affinity for BoNT/A2 while maintaining high affinity binding to BoNT/A1. Thermodynamics of antiody binding to BoNT/A1 and BoNT/A2 were studied and the crsytal structure of the antibody complexed with BoNT/A1 was solved.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
