CloneID: CR1
Heavy Chain modification: Fc Silent™
Antigen Long Description: 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).
Buffer Composition: PBS with 0.02% Proclin 300.
Chimeric Use Statement: This chimeric mouse antibody was made using the variable domain sequences of the original Human IgG1 format, for improved compatibility with existing reagents, assays and techniques.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: P10845
Specificity Statement: 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.
Application Notes (Clone): 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.