Anti-Rituximab Antibodies from ACROBiosystems

Anti-Rituximab Antibody Information

Source: Mouse monoclonal antibody is produced from a hybridoma resulting from fusion of SP2/0 myeloma and B-lymphocytes obtained from a mouse immunized with Rituximab F(ab')2.

Isotype: IgG1/kappa

Specificity: Recognizes Rituximab specifically, no cross reactivity with other humanized antibodies.

Purity: >95% as determined by SDS-PAGE.

Formulation: Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.

Reconstitution: Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

Storage

For long term storage, the product should be stored at lyophilized state at -20 °C or lower.

Please avoid repeated freeze-thaw cycles.

No activity loss was observed after storage at:

  • 4-8 °C for 12 months in lyophilized state;
  • -70 °C for 3 years under sterile conditions after reconstitution.

SDS-PAGE

Anti-Rituximab Antibodies on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA

Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized rituximab at 5 µg/ml, add increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y35, 10% human serum) and then add biotinylated rituximab at 5 µg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 20 ng/mL.

Demonstration of the specificity of Anti-Rituximab Antibodies (Cat. No. RIB-Y35) to the rituximab.

Detection of rituximab by anti-idiotypic capture ELISA in serum. Immobilized Anti-Rituximab Antibodies (Cat. No. RIB-Y35) at 0.5 μg/mL (100 μL/well), add increasing concentrations of rituximab (0.1% human serum). Detection was performed using goat anti-human IgG with a sensitivity of 0.156 μg/mL.

Bioactivity-SPR

Anti-Rituximab Antibodies (mouse IgG1, Cat. No. RIB-Y35) captured on CM5 chip via anti-mouse antibodies surface, can bind Human rituximab with an affinity constant of 0.03 nM.

Bioactivity-FACS

FACS analysis shows that the binding of rituximab to 293F overexpressing CD20 was inhibited by increasing concentration of Anti-Rituximab Antibodies (Cat. No. RIB-Y35). The concentration of rituximab used is 10 ng/ml. The IC50 is 0.013 μg/ml (Routinely tested).

Background

Rituxan is a genetically engineered chimeric murine/human monoclonal antibody directed against the CD20 antigen found on the surface of normal and malignant B lymphocytes. The antibody is an IgG1 kappa immunoglobulin containing murine light- and heavy-chain variable region sequences and human constant region sequences. Rituximab is composed of two heavy chains of 451 amino acids and two light chains of 213 amino acids