Infliximab antibody | AbD19376
To confirm antibody specificity, a microtiter plate was coated overnight with various antigens at a concentration of 5μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HCA214 at 2μg/ml, followed by Mouse Anti-Histidine-tag:HRP Antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with Human IgG1/kappa, Human IgG1/lambda or infliximab at a concentration of 5μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HCA214 titrated to the given concentrations in PBST+10% Human serum, followed by Mouse Anti-Histidine-tag:HRP Antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
To assess antibody sensitivity, a microtiter plate was coated overnight with HCA214 at 5μg/ml. After washing and blocking with PBST+5% BSA, infliximab was added at the given concentrations. Detection was performed using Mouse Anti-Human IgG (Fc) CH2 domain:HRP Antibody (MCA647P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
HCA214 does not inhibit the binding of infliximab to TNF-α
To assess the ability of HCA214 to inhibit the binding of infliximab to its target molecule, a microtiter plate was coated overnight with TNF-α at a concentration of 1µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of HCA214 titrated into a constant amount of infliximab (IFX, 0.3μg/ml) was added. Free infliximab, still capable of binding to the plate, was detected using Mouse Anti-Human IgG (Fc) CH2 domain:HRP Antibody (MCA647P).
Infliximab bound to TNF-α still binds to HCA214
HCA214 was coated overnight onto a microtiter plate at a concentration of 5µg/ml. After washing and blocking with PBST+5% BSA, infliximab alone (green triangles), with 0.5 fold molar excess of TNF-α (red circles) or with 5.0 fold molar excess of TNF-α (black squares) was added. Free or TNF-α bound infliximab binding to HCA214 was detected using Mouse Anti-Human IgG (Fc) CH2 domain:HRP Antibody (MCA647P).
A microtiter plate was coated overnight with HCA214 at a concentration of 5μg/ml (black circles) or 1μg/ml (blue squares). After washing and blocking with PBST+5% BSA, infliximab spiked in 10% Human serum was added at the given concentrations. Detection was performed using HRP-conjugated Antibodies HCA213 (black circles) or HCA216 (blue squares).
Human anti Infliximab
Anti-infliximab antibody is a non-neutralizing anti-idiotypic monovalent Fab antibody; it is ideal for bioanalytical assays for infliximab and biosimilars. It does not inhibit the binding of infliximab to TNF alpha and so detects total drug. It is recommended as PK bridging ELISA capture antibody.
- Product Type
- Monoclonal Antibody
- Clone
- AbD19376
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA214 | E | 0.1 mg |
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| HCA214G | E | 1 mg |
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Human Anti-Infliximab Antibody, clone AbD19376 is an anti-idiotypic antibody that specifically recognizes the infliximab monoclonal antibody and does not inhibit the binding of infliximab to its target, tumor necrosis factor alpha (TNFα). It can be used in bioanalytical assays to measure the levels of infliximab and biosimilar products, such as Inflectra and Renflexis, in patient samples.
Clone AbD19376 recognizes both the free infliximab monoclonal antibody and infliximab when bound to TNFα (the drug/target complex); it is not paratope-specific and it is non-inhibitory. This antibody can be used as a capture antibody in a pharmacokinetic (PK) bridging ELISA to detect total drug (free, partially bound and fully bound) with HCA216P as the detection antibody.
Infliximab (branded as Remicade) is a chimeric monoclonal antibody drug (IgG1/kappa) that has been approved for treatment of psoriasis, Crohn's disease, ankylosing spondylitis, psoriatic arthritis, rheumatoid arthritis, and ulcerative colitis. It acts by blocking the binding of TNFα to its receptors. Infliximab also induces apoptosis of TNFα expressing T-lymphocytes.
View a summary of all Anti-Infliximab Antibodies.
Clone AbD19376 recognizes both the free infliximab monoclonal antibody and infliximab when bound to TNFα (the drug/target complex); it is not paratope-specific and it is non-inhibitory. This antibody can be used as a capture antibody in a pharmacokinetic (PK) bridging ELISA to detect total drug (free, partially bound and fully bound) with HCA216P as the detection antibody.
Infliximab (branded as Remicade) is a chimeric monoclonal antibody drug (IgG1/kappa) that has been approved for treatment of psoriasis, Crohn's disease, ankylosing spondylitis, psoriatic arthritis, rheumatoid arthritis, and ulcerative colitis. It acts by blocking the binding of TNFα to its receptors. Infliximab also induces apoptosis of TNFα expressing T-lymphocytes.
View a summary of all Anti-Infliximab 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.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Infliximab
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=0.7 nM by real time, label-free molecular interaction analysis on immobilized infliximab.
- 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.
Remicade is a registered trademark of Janssen Biotech Inc. Inflectra is a registered trademark of Hospira, a Pfizer Company. RENFLEXIS is a trademark of Merck Sharp & Dohme Corp.
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 Infliximab 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 as a capture antibody in an Infliximab bridging ELISA together HRP-conjugated HCA213 or HCA216 as the detection reagent.
Protocol: PK bridging ELISA to measure total 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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| 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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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Goat anti Histidine Tag:Biotin | AHP1656B | E WB | 0.1 mg |
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| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Infliximab | HCA213 | E | 0.1 mg |
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| Human anti Infliximab | HCA215 | E | 0.1 mg |
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| Human anti Infliximab | HCA216 | E | 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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| 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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