Cetuximab antibody | AbD19376_hIgG1
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP conjugated Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated over night with human IgG1/kappa, human IgG1/lambda, rituximab or cetuximab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) in a monovalent format, titrated to the given concentrations in PBST+10% human serum, followed by Mouse Anti-Penta Histidine Tag:HRP (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated over night with Human Anti-Cetuximab Antibody, clone AbD19834 (HCA220) 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 cetuximab. Detection was performed using HRP conjugated Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated over night with epidermal growth factor receptor (EGFR) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of cetuximab (CTX, 0.3 μg/ml) plus increasing concentrations of the non-inhibitory Human Anti-Cetuximab Antibody (HCA228) was added. In parallel experiments, HCA228 was replaced with inhibitory Human Anti-Cetuximab Antibodies of varying affinity (HCA220, HCA221 or HCA223). For this assay, HCA221, HCA223 and HCA228 were used in their monovalent Fab format. Free cetuximab, still capable of binding to the EGFR-coated plate, was detected using Goat Anti-Human IgG F(ab')2:HRP (STAR126P).
Filter by Application:
E ResetHuman anti Cetuximab
- Product Type
- Monoclonal Antibody
- Clone
- AbD19376_hIgG1
- Isotype
- IgG1
- Specificity
- Cetuximab
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| Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1, is a non-inhibitory anti-idiotypic antibody that binds to cetuximab. This antibody recognizes an epitope outside the cetuximab binding site and so does not inhibit the binding of the drug to its target, epidermal growth factor receptor (EGFR). This epitope is shared with other chimeric antibodies that are highly homologous to cetuximab including infliximab but excluding rituximab. The antibody is suitable for the development of bioanalytical assays for cetuximab and biosimilars. Clone AbD19376_hIgG1 is a fully human recombinant monoclonal with IgG1 isotype and is suitable as a surrogate positive control or calibrator in an anti-drug antibody (ADA) assay. In addition, it is suitable as a detection antibody in pharmacokinetic (PK) bridging ELISA, with Anti-Cetuximab Antibody HCA220 as the capture antibody. This combination enables measurement of free and partially bound cetuximab, which is equivalent to total drug when trough serum levels are above 4 μg/ml. Cetuximab (brand name Erbitux) is a chimeric monoclonal antibody drug (IgG1/kappa) approved for treatment of KRAS wild-type metastatic colorectal cancers and squamous cell carcinoma of the head and neck. This therapeutic antibody is directed against EGFR and inhibits tumor cell proliferation by blocking the interaction of epidermal growth factor (EGF) with its receptor. View a summary of all anti cetuximab antibodies. |
- Product Form
- Human IgG1 antibody selected from the HuCAL? phage display library and expressed in a human cell line. This antibody is supplied as a liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Chimeric monoclonal antibody
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=39 nM by real time, label-free molecular interaction analysis on immobilized cetuximab.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- 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.
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.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| 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 direct ELISA or when conjugated to HRP, as a detection antibody in a cetuximab bridging ELISA for PK assay development together with HCA220 as the capture reagent.
Protocol: PK bridging ELISA to measure total drug
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Cetuximab | HCA220 | E | 0.1 mg |
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| Description | Human anti Cetuximab | ||||||
| Human anti Cetuximab | HCA221 | E | 0.1 mg |
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| List Price | Your Price | ||||||
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| Description | Human anti Cetuximab | ||||||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | Log in | |||
| List Price | Your Price | ||||||
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| Description | LYNX Rapid HRP Antibody Conjugation Kit | ||||||
References for Cetuximab antibody
-
Bose, M.E. et al. (2016) Simulated Respiratory Secretion for Use in the Development of Influenza Diagnostic Assays.
PLoS One. 11 (11): e0166800. -
Kashiwagi, N. et al. (2017) Method for measuring anti-drug antibody
US Patent Application US20170315118A1
Further Reading
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Gomez, D. et al. (2013) Cetuximab therapy in the treatment of metastatic colorectal cancer: the future frontier?
Int J Surg. 11 (7): 507-13.
- 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.
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