Adalimumab antibody | AbD18654
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, Human Anti-Adalimumab Antibody, clone AbD18654 (HCA202) was added at a concentration of 2 μg/ml. Detection was performed using Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with Human Anti-Adalimumab Antibody, AbD18654 (HCA202) 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 adalimumab. Detection was performed using Human Anti-Adalimumab:HRP Antibody, clone AbD18655_hIgG1 (HCA204P) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with adalimumab (black), infliximab (red), alemtuzumab (green), human IgG1/kappa (blue), BSA (light blue) at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, Human Anti-Adalimumab Antibody, clone AbD18654 (HCA202) was added . Detection was performed using Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with Recombinant Human TNFα (PHP051) at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, a pre-incubated mixture of adalimumab (0.3 μg/ml), spiked with increasing concentrations of Human Anti-Adalimumab Antibody, clone AbD18654 (HCA202) was added. Free adalimumab still capable of binding to the Human Recombinant TNFα coated plate, was detected using Goat Anti-Human IgG F(ab’)2:HRP (STAR126P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti Adalimumab
Anti-adalimumab antibody is a recombinant, neutralizing anti-idiotypic antibody in monovalent Fab format; it is ideal for bioanalytical assay development to measure levels of adalimumab or biosimilars. It is recommended as capture antibody in PK bridging ELISA with HCA204/HCA204P.
- Product Type
- Monoclonal Antibody
- Clone
- AbD18654
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA202 | E | 0.1 mg |
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| HCA202G | E | 1 mg |
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Human Anti-Adalimumab Antibody, clone AbD18654, is a high affinity anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug adalimumab and inhibits binding to its target, TNFα. The antibody can be used to measure the levels of adalimumab and biosimilar products in bioanalytical assays.
Clone AbD18654 is recommended as a capture antibody in a pharmacokinetic (PK) bridging assay when used in combination with HRP-conjugated detection antibody (HCA204P). The assay will measure free drug, as both antibodies are inhibitory.
Adalimumab (branded as Humira) is a fully human monoclonal antibody (IgG1/kappa) approved for the treatment of ankylosing spondylitis, Crohn’s disease, psoriasis and several forms of arthritis. The antibody is specific for TNFα, and acts by blocking the activation of TNF receptors by TNFα. This inactivation down-regulates the inflammatory reactions associated with autoimmune diseases.
View a summary of all Anti-Adalimumab Antibodies.
Clone AbD18654 is recommended as a capture antibody in a pharmacokinetic (PK) bridging assay when used in combination with HRP-conjugated detection antibody (HCA204P). The assay will measure free drug, as both antibodies are inhibitory.
Adalimumab (branded as Humira) is a fully human monoclonal antibody (IgG1/kappa) approved for the treatment of ankylosing spondylitis, Crohn’s disease, psoriasis and several forms of arthritis. The antibody is specific for TNFα, and acts by blocking the activation of TNF receptors by TNFα. This inactivation down-regulates the inflammatory reactions associated with autoimmune diseases.
View a summary of all Anti-Adalimumab Antibodies.
Product Details
- Product Form
- A monovalent human recombinant Fab (lambda 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
- Adalimumab
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=0.15 nM by real time, label-free molecular interaction analysis on immobilized adalimumab.
- 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
- Sold under license of U.S. Patents 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad.
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 or in an ELISA bridging assay as a capture antibody together with HCA204P as the detection reagent.
Protocol: PK bridging ELISA to measure free drug
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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 | HCA204 | E | 0.1 mg |
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| Human anti Adalimumab:HRP | HCA204P | E | 0.1 mg |
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| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | C* E | 0.2 mg |
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| Recombinant Human TNF Alpha | PHP051 | E FN WB | 50 µg |
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| Goat anti Human IgG F(ab')2:HRP | STAR126P | E WB | 0.5 ml |
|
Product Specific References
References for Adalimumab antibody
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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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