Rituximab antibody | MB2A4
A microtiter plate was coated overnight with Human Anti-Rituximab Antibody, clone AbD18423 (HCA186) at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, 10% Human Serum was added spiked with increasing concentrations of rituximab. Detection was performed using Rat Anti-Rituximab:HRP Antibody, clone MB2A4 (MCA2260P) at a concentration of 3 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three independent measurements.
Figure A. PE conjugated rituximab pre-incubated for 1h with Rat Anti-Rituximab Antibody, clone MB2A4 (MCA2260) (red) or Rat IgG2a isotype control (MCA6005) (grey) in ratio 1:1. Figure B. PE conjugated rituximab pre-incubated for 1h with Rat Anti-Rituximab Antibody, clone MB2A4 (MCA2260) (red) or Rat IgG2a isotype control (MCA6005) (grey) in ratio 5:1 (Anti-Rituximab or isotype:rituximab). All experiments performed on Daudi cell line expressing CD20 and incubated for 30 min with the antibody mix.
A microtiter plate was coated overnight with ocrelizumab, rituximab, infliximab and human IgG1 kappa at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Rat Anti-Rituximab:HRP Antibody, clone MB2 A4 (MCA2260P) titrated to the given concentrations in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Rat anti Rituximab
- Product Type
- Monoclonal Antibody
- Clone
- MB2A4
- Isotype
- IgG2a
- Specificity
- Rituximab
Antibody Quality and Performance Guarantee
| Rat Anti-Rituximab Antibody, clone MB2A4, is an anti-idiotypic antibody that recognizes the monoclonal antibody drug rituximab. The antibody can be used to measure the levels of rituximab and biosimilar products in bioanalytical assays. Rat Anti-Rituximab Antibody also recognizes ocrelizumab, a second-generation humanized anti-CD20 antibody that binds to an epitope on CD20 that is identical or overlapping with the rituximab epitope. Clone MB2A4 has been used in ELISA to monitor the levels of rituximab in patient serum following therapy (Cragg et al. 2004 and Hampson et al. 2010). Clone MB2A4 has been used to detect rituximab bound to the surface of the Raji B cell line, however detection of rituximab bound in vivo to B-CLL cells has not been demonstrated. It is possible that complement deposition on rituximab opsonised cells inhibits binding of the Anti-Rituximab Antibody to cell bound rituximab (Beum et al. 2004). Inhibition experiments carried out with Daudi cells demonstrated that this antibody is inhibitory at a ratio of 5:1 antibody:rituximab, but does not inhibit rituximab binding to CD20 at a ratio of 1:1. Rituximab (reference product branded as Rituxan) is a chimeric mouse/human monoclonal antibody approved for the treatment of certain autoimmune diseases and cancer, including non-Hodgkin's lymphoma, chronic lymphocytic leukemia and rheumatoid arthritis. The antibody is specific for the cell surface protein CD20, which is widely expressed on B cells. Through three different mechanisms of action it eliminates B cells from the body, enabling the development of a new population of healthy B cells. View a summary of all Anti-Rituximab Antibodies. |
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Buffer Solution
- TRIS buffered saline
- Preservative Stabilisers
- MCA2260: <0.1% sodium azide (NaN3)
- MCA2260G: < 0.1% Sodium Azide (NaN3)
- Immunogen
- F(ab)2 fragment of Rituximab
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised rats were fused with cells of the NS-1 mouse myeloma cell line
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- Rituxan is a trademark of Biogen Idec/Genentech in the USA.
MabThera is a trademark of Roche in Europe.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
||
| Flow Cytometry | |
50 ug/ml |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10μl of the suggested working dilution to label 106 cells in 100μl
- ELISA
- Rat anti Rituximab antibody, clone MB2A4 may be used in a direct ELISA or as a detection reagent, when conjugated to HRP, in a sandwich ELISA together with HCA186 as the capture reagent. Protocol: PK bridging ELISA to measure free drug
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| Description | Goat F(ab')2 anti Rat IgG:RPE (Mouse Adsorbed) | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Human anti Rituximab | HCA061 | E | 0.1 mg |
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| Description | Human anti Rituximab | ||||||
| Human anti Rituximab | HCA186 | E | 0.1 mg |
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| Description | Human anti Rituximab | ||||||
References for Rituximab antibody
-
Cragg, M. S. et al. (2004) A new anti-idiotype antibody capable of binding rituximab on the surface of lymphoma cells.
Blood. 104:2540-2 -
Cragg, M.S. et al. (2004) Apparent modulation of CD20 by rituximab: an alternative explanation.
Blood. 103 (10): 3989-90; author reply 3990-1. -
Pers, J.O. et al. (2007) BAFF-modulated repopulation of B lymphocytes in the blood and salivary glands of rituximab-treated patients with Sjögren's syndrome.
Arthritis Rheum. 56: 1464-77. -
Hampson, G. et al. (2010) Validation of an ELISA for the determination of rituximab pharmacokinetics in clinical trials subjects.
J Immunol Methods. 360 (1-2): 30-8. -
Blasco, H. et al. (2007) Evaluation of a peptide ELISA for the detection of rituximab in serum.
J Immunol Methods.325: 127-39. -
Daydé, D. et al. (2009) Tumor burden influences exposure and response to rituximab: pharmacokinetic-pharmacodynamic modeling using a syngeneic bioluminescent murine model expressing human CD20.
Blood. 113: 3765-72. -
Aung, T. et al. (2011) Exosomal evasion of humoral immunotherapy in aggressive B-cell lymphoma modulated by ATP-binding cassette transporter A3.
Proc Natl Acad Sci U S A. 108: 15336-41. -
Schmidt, E. et al. (2009) Immunogenicity of rituximab in patients with severe pemphigus.
Clin Immunol. 132: 334-41.
View The Latest Product References
-
McDonald, V. et al. (2010) Rituximab pharmacokinetics during the management of acute idiopathic thrombotic thrombocytopenic purpura.
J Thromb Haemost. 8: 1201-8. -
Kagan, L. et al. (2012) Subcutaneous Absorption of Monoclonal Antibodies: Role of Dose, Site of Injection, and Injection Volume on Rituximab Pharmacokinetics in Rats.
Pharm Res. 29: 490-499 -
Kagan, L. and Mager, D.E. (2013) Mechanisms of subcutaneous absorption of rituximab in rats.
Drug Metab Dispos. 41: 248-55. -
Liu, X.F. et al. (2012) Validation of a Gyrolab™ assay for quantification of rituximab in human serum.
J Pharmacol Toxicol Methods. 65: 107-14. -
Kagan, L. et al. (2014) Interspecies pharmacokinetic modeling of subcutaneous absorption of rituximab in mice and rats.
Pharm Res. 31: 3265-73. -
Blasco, H. et al. (2009) Pharmacokinetics of rituximab associated with CHOP chemotherapy in B-cell non-Hodgkin lymphoma.
Fundam Clin Pharmacol. 23: 601-8. -
Illidge, T.M. et al. (2016) Short duration immunochemotherapy followed by radioimmunotherapy consolidation is effective and well tolerated in relapsed follicular lymphoma: 5-year results from a UK National Cancer Research Institute Lymphoma Group study.
Br J Haematol. 173 (2): 274-82. -
Vacher, P. et al. (2015) Localized Store-Operated Calcium Influx Represses CD95-Dependent Apoptotic Effects of Rituximab in Non-Hodgkin B Lymphomas.
J Immunol. pii: 1402942. -
Komori, M. et al. (2016) Insufficient disease inhibition by intrathecal rituximab in progressive multiple sclerosis.
Ann Clin Transl Neurol. 3 (3): 166-79. -
Kashiwagi, N. et al. (2017) Method for measuring anti-drug antibody
US Patent Application US20170315118A1 -
Lioger, B. et al. (2017) Antigenic burden and serum IgG concentrations influence rituximab pharmacokinetics in rheumatoid arthritis patients.
Br J Clin Pharmacol. 83 (8): 1773-81. -
Zhang, Y. et al. (2013) Stability of stock and diluted rituximab.
Am J Health Syst Pharm. 70 (5): 436-8. -
Desbois, A.C. et al. (2020) Rituximab-associated Vasculitis Flare: Incidence, Predictors, and Outcome.
J Rheumatol. 47 (6): 896-902.
- Synonyms
- Mabthera
- Rituxan
- RRID
- AB_323787
View more products with RITUXIMAB specificity
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