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 | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA248 | E | 0.1 mg |
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| HCA248G | E | 1 mg |
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Human Anti-Natalizumab Antibody, clone AbD21375 is a paratope specific, anti-idiotypic antibody that specifically recognizes free natalizumab but not free human alpha-4/beta-1 integrin or the drug/integrin complex. It inhibits the binding of natalizumab to its target and can be used to measure levels of free natalizumab and biosimilar products in bioanalytical and patient monitoring assays.
A pair of anti-natalizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as a capture antibody, paired with an HRP conjugated Anti-Natalizumab Antibody in full immunoglobulin format, clone AbD21375_hlgG1 (HCA249) as the detection antibody.
Natalizumab (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. It is directed against alpha-4 integrin (CD49d), and binds to alpha-4/beta-1 integrin on the surface of circulating leucocytes, except neutrophils. 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.
View a summary of all anti-natalizumab antibodies
A pair of anti-natalizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as a capture antibody, paired with an HRP conjugated Anti-Natalizumab Antibody in full immunoglobulin format, clone AbD21375_hlgG1 (HCA249) as the detection antibody.
Natalizumab (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. It is directed against alpha-4 integrin (CD49d), and binds to alpha-4/beta-1 integrin on the surface of circulating leucocytes, except neutrophils. 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.
View a summary of all anti-natalizumab antibodies
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
- Pack Size: 0.1 mgAntibody concentration 0.5 mg/mlPack Size: 1 mgAntibody concentration 1.0 mg/ml
Storage Information
- Storage
- 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. - Guarantee
- 12 months from date of despatch
More Information
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
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
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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Natalizumab | HCA249 | E | 0.1 mg |
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| Human anti Natalizumab | HCA250 | E | 0.1 mg |
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| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | ||
| Rat anti DYKDDDDK Tag:HRP | MCA4764P | E P WB | 0.1 mg |
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| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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