Natalizumab antibody | AbD21375
A microtiter plate was coated over night with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti natalizumab (clone AbD21375) at a concentration of 2 μg/ml followed by Rat anti DYKDDDDK Tag:HRP (MCA4764P) in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate
A microtiter plate was coated over night with natalizumab, human IgG4/kappa, human IgG1/kappa, human IgG1/lambda, tocilizumab or omalizumab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti natalizumab (clone AbD21375) titrated to the given concentrations in PBST, followed by an HRP conjugated anti-His tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements
A microtiter plate was coated over night with Human anti natalizumab (clone AbD21375) 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 natalizumab. Detection was performed using HRP-conjugated Human anti natalizumab (clone AbD21375) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a Lynx Rapid HRP Antibody Conjugation Kit®. Data is presented as the mean of three measurements
A microtiter plate was coated over night with Human anti natalizumab (clone AbD21375) 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 natalizumab. Detection was performed using HRP-conjugated Human anti natalizumab (clone AbD21375_hIgG1, HCA249) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®. Data is presented as the mean of three measurements
A microtiter plate was coated over night with alpha-4/beta-1 integrin at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of natalizumab (0.3 μg/ml) plus increasing concentrations of Human anti natalizumab (clone AbD21375) was added. Free natalizumab, still capable of binding to the alpha-4/beta-1 integrin coated plate, was detected using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements
Human anti Natalizumab
Anti-natalizumab antibody is a recombinant, neutralizing anti-idiotypic antibody in monovalent Fab format; it is ideal for bioanalytical assay development to measure levels of natalizumab or biosimilars. It is recommended as capture antibody in PK bridging ELISA with HCA249/HCA249P.
- Product Type
- Monoclonal Antibody
- Clone
- AbD21375
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA248 | E | 0.1 mg |
Human anti natalizumab, clone AbD21375 is a Type 1 anti-idiotypic antibody that specifically recognizes free natalizumab but not free human alpha-4/beta-1 integrin or the drug/integrin complex. Clone AbD21375 inhibits the binding of natalizumab to alpha-4/beta-1 integrin and can be used to measure free natalizumab levels in serum from patients. This clone is also suitable for pharmacokinetic (PK) assay development.
Natalizumab, marketed under the brand name Tysabri®, is a humanized IgG4/kappa monoclonal antibody approved as a monotherapy for the treatment of relapsing forms of multiple sclerosis and for the treatment of Crohn’s disease. Natalizumab is also being evaluated for potential clinical use in the treatment of B-cell malignancies. This therapeutic antibody, directed against alpha-4 integrin (CD49d), binds to alpha-4/beta-1 integrin on the surface of circulating leucocytes. It is believed to act by blocking the interaction of alpha-4/beta-1 integrin with vascular cell adhesion molecule-1 (VCAM-1), preventing the adherence and transport of inflammatory immune cells across the intestinal lining and blood-brain barrier.
Clone AbD21375 is one of a range of Type 1 (HCA249, HCA250), anti-idiotypic antibodies to natalizumab available from Bio-Rad and forms part of a wider and expanding portfolio of anti-idiotypic antibodies that have been developed for the detection of biotherapeutics.
Natalizumab, marketed under the brand name Tysabri®, is a humanized IgG4/kappa monoclonal antibody approved as a monotherapy for the treatment of relapsing forms of multiple sclerosis and for the treatment of Crohn’s disease. Natalizumab is also being evaluated for potential clinical use in the treatment of B-cell malignancies. This therapeutic antibody, directed against alpha-4 integrin (CD49d), binds to alpha-4/beta-1 integrin on the surface of circulating leucocytes. It is believed to act by blocking the interaction of alpha-4/beta-1 integrin with vascular cell adhesion molecule-1 (VCAM-1), preventing the adherence and transport of inflammatory immune cells across the intestinal lining and blood-brain barrier.
Clone AbD21375 is one of a range of Type 1 (HCA249, HCA250), anti-idiotypic antibodies to natalizumab available from Bio-Rad and forms part of a wider and expanding portfolio of anti-idiotypic antibodies that have been developed for the detection of biotherapeutics.
Product Details
- Product Form
- A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in E. coli. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Natalizumab
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD = 2.1 nM by real time, label free molecular interaction analysis on immobilized natalizumab.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use - Shelf Life
- 12 months from date of despatch
More Information
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany. Tysabri® is a registered trademark Biogen Idec and Elan.
His-tag is a registered trademark of EMD Biosciences. - Licensed Use
- For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
- Regulatory
- For research purposes only
Applications of Natalizumab antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |
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.
- Technical Advice
- Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- This product may be used in a either a direct ELISA or indirect ELISA both as a capture reagent and as a detection reagent.
Protocol: PK bridging ELISA to measure free drug
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | 50 ml | E IY | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | 1 Conjugation For 400µg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | 3 Conjugations For 400µg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK003P | 1 Conjugation For 4mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK004P | 5 Conjugations For 4mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK005P | 1 Conjugation For 20mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK006P | 3 Conjugations For 40µg Antibody | CJ | ||
| Rat anti DYKDDDDK Tag:HRP | MCA4764P | 0.1 mg | E P WB | ||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | 125 µl | E WB | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | 0.2 mg | C* E |
