Adalimumab antibody | AbD20350
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 adalimumab (clone 20350) at a concentration of 2 μg/ml, followed by an HRP-conjugated mouse anti His tag antibody in HISPEC assay diluent (BUF049A) and a fluorogenic peroxidase substrate.
A microtiter plate was coated over night with human TNFα at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of adalimumab. Detection was performed using unlabeled (red line) or HRP-conjugated (black line) Human anti adalimumab (clone 20350) at a concentration of 2 μg/ml in PBST. Unlabeled anti adalimumab was detected by an HRP-conjugated mouse anti His tag antibody in HISPEC assay diluent (BUF049C) and a fluorogenic peroxidase substrate. HRP-conjugated anti adalimumab was detected directly with fluorogenic substrate. HRP-conjugation of the antibody used in this assay was performed using a Lynx Rapid HRP Antibody Conjugation Kit (LNK001P).
A microtiter plate was coated overnight with Recombinant Human TNFα (PHP051), adalimumab, infliximab and Human IgG1 kappa control at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, the human TNFα/adalimumab and TNFα/infliximab complexes were formed by adding 2 μg/ml adalimumab or infliximab to the wells coated with TNFα. Detection was performed using Human Anti-Adalimumab Antibody, clone AbD20350 (HCA231) titrated to the given concentrations in PBST followed by Mouse Anti-Penta Histidine Tag:HRP (MCA5995P) in HISPEC Assay Diluent (BUF049A) and a fluorogenic peroxidase substrate.
Human anti Adalimumab (Drug/Target Complex)
Anti-adalimumab drug-target complex antibody specifically recognizes the adalimumab-TNF alpha complex exclusively and detects adalimumab or biosimilars only when bound to TNF alpha. This affinity-matured antibody is in monovalent Fab format and is suitable for a PK antigen capture format assay.
- Product Type
- Monoclonal Antibody
- Clone
- AbD20350
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA231 | E | 0.1 mg |
|
Human anti Adalimumab (drug/target complex), clone AbD20350, is an affinity matured, Type 3 anti-idiotypic antibody that specifically recognizes adalimumab complexed with TNFα. It does not recognize the free drug or unbound TNFα. Clone AbD20350 can be used in assays to measure adalimumab that has been captured via immobilized TNFα, thereby avoiding the bridging assay format. It can also be used to measure the level of the drug/target complex and biosimilar products using bioanalytical assays.
Adalimumab (brand name Humira®) is a human monoclonal antibody (IgG1/kappa) approved for the treatment of rheumatoid, psoriatic and juvenile idiopathic arthritis, ankylosing spondylitis, Crohn’s disease and ulcerative colitis. This therapeutic antibody is directed against the pro-inflammatory cytokine tumor necrosis factor alpha (TNFα), blocking its interaction with the p55 (TNFR1, CD120a) and p75 (TNFR2, CD120b) cell surface TNF receptors, resulting in a down-regulation of the abnormal inflammatory response that is characteristic of autoimmune disease.
View a summary of all anti-adalimumab antibodies.
Adalimumab (brand name Humira®) is a human monoclonal antibody (IgG1/kappa) approved for the treatment of rheumatoid, psoriatic and juvenile idiopathic arthritis, ankylosing spondylitis, Crohn’s disease and ulcerative colitis. This therapeutic antibody is directed against the pro-inflammatory cytokine tumor necrosis factor alpha (TNFα), blocking its interaction with the p55 (TNFR1, CD120a) and p75 (TNFR2, CD120b) cell surface TNF receptors, resulting in a down-regulation of the abnormal inflammatory response that is characteristic of autoimmune disease.
View a summary of all anti-adalimumab antibodies.
Product Details
- Product Form
- A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in E. coli. This antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This product is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Adalimumab.
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=3.2 nM by real time, label-free molecular interaction analysis on immobilized adalimumab/TNFα complex.
- Approx. Protein Concentrations
- Antibody concentration 0.5 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.
Humira® is a registered trademark of Abbott Laboratories.
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 Adalimumab 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 an indirect ELISA to detect the adalimumab/TNFα complex or to detect adalimumab bound to immobilized TNFα.
Protocol: PK antigen capture ELISA to measure bound drug exclusively
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Secondary Antibodies Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Rabbit anti DYKDDDDK Tag:HRP | AHP1074P | E WB | 0.1 mg |
|
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| Goat anti Histidine Tag:Biotin | AHP1656B | E WB | 0.1 mg |
|
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| Mouse anti Histidine Tag:Alk.Phos. | MCA1396A | E WB | 0.1 mg |
|
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| Mouse anti Histidine Tag:Biotin | MCA1396B | E WB | 0.1 mg |
|
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| 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 |
|
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Adalimumab (Drug/Target Complex) | HCA232 | E | 0.1 mg |
|
|
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | ||
| Recombinant Human TNF Alpha | PHP051 | E FN WB | 50 µg |
|
Product Specific References
References for Adalimumab antibody
-
Harth, S. et al. (2019) Generation by phage display and characterization of drug-target complex-specific antibodies for pharmacokinetic analysis of biotherapeutics.
MAbs. 11 (1): 178-190.
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