Mouse monoclonal antibody is produced from a hybridoma resulting from fusion of SP2/0 myeloma and B-lymphocytes obtained from a mouse immunized with Bevacizumab F(ab')2.
Recognizes Bevacizumab specifically, no cross reactivity with other humanized antibodies.
The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A specially optimized long-spacer biotin reagent (13.5 angstroms) is used in this product to minimize potential steric hindrance.
The biotin to protein ratio is 1-3 as determined by the HABA assay.
>90% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 years under sterile conditions after reconstitution.
Biotinylated Anti-Bevacizumab Antibodies (recommended for PK/PD) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Detection of bevacizumab by bridging ELISA in serum. Immobilized Anti-Bevacizumab Antibodies (recommended for PK/PD) (Cat. No. BEB-Y10) at 2 μg/mL, add increasing concentrations of bevacizumab (10% human serum) and then add Biotinylated Anti-Bevacizumab Antibodies (recommended for PK/PD) (Cat. No. BEB-BY13) at 2 μg/mL. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 0.4 μg/mL.
Immobilized bevacizumab at 1 μg/mL (100 μL/well) can bind Biotinylated Anti-Bevacizumab Antibodies (recommended for PK/PD) (Cat. No. BEB-BY13) with a linear range of 0.78-25.5 ng/mL.
ELISA analysis shows that the binding of bevacizumab to ActiveMax® Human VEGF165, Tag Free (HPLC-verified) (Cat. No. VE5-H4210) was inhibited by increasing concentration of Biotinylated Anti-Bevacizumab Antibodies (recommended for PK/PD) (Cat. No. BEB-BY13). The concentration of bevacizumab used is 4 ng/mL.
A recombinant humanized monoclonal IgG1 antibody that binds to and inhibits the biologic activity of human vascular endothelial growth factor (VEGF). Bevacizumab contains human framework regions and the complementarity-determining regions of a murine antibody that binds to VEGF. Bevacizumab is produced in a Chinese Hamster Ovary mammalian cell expression system in a nutrient medium containing the antibiotic gentamicin and has a molecular weight of approximately 149 kilodaltons.