Cetuximab antibody | AbD19834
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-Cetuximab Antibody, clone AbD19834 (HCA220) at a concentration of 2 μg/ml, 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 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 AbD19834 (HCA220) 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 1 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 AbD19830_hIgG1 (HCA221P) 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 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 the non-inhibitory Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228P) at a concentration of 2 μg/ml 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 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 Human Anti-Cetuximab Antibody HCA220, HCA221 or HCA223, or the non-inhibitory antibody HCA228, was added. For this assay HCA221, HCA223 and HCA228 were used in the 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).
Human anti Cetuximab
Anti-Cetuximab Antibody is a recombinant, inhibitory, anti-idiotypic antibody in monovalent Fab format; it is ideal for bioanalytical assay development for cetuximab and biosimilars. It is suitable as a capture antibody in PK bridging ELISA format with HCA221 for an assay to detect free drug.
- Product Type
- Monoclonal Antibody
- Clone
- AbD19834
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA220 | E | 0.1 mg |
|
Human Anti-Cetuximab Antibody, clone AbD19834, is an anti-idiotypic antibody that binds specifically to cetuximab and inhibits the binding of the drug to its target, epidermal growth factor receptor (EGFR). It can be used in bioanalytical assays to measure the levels of cetuximab and biosimilar products in preclinical research and clinical trials.
Clone AbD19834 is recommended as the capture antibody in a pharmacokinetic (PK) bridging ELISA. When paired with another inhibitory antibody, HRP conjugated Anti-Cetuximab Antibody clone AbD19830_hIgG1 (HCA221P), as detection antibody, free drug can be measured. When paired with the non-inhibitory Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228P), free and partially bound drug can be measured, 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.
Clone AbD19834 is recommended as the capture antibody in a pharmacokinetic (PK) bridging ELISA. When paired with another inhibitory antibody, HRP conjugated Anti-Cetuximab Antibody clone AbD19830_hIgG1 (HCA221P), as detection antibody, free drug can be measured. When paired with the non-inhibitory Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228P), free and partially bound drug can be measured, 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 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
- Cetuximab.
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD = 0.5 nM by real time, label free molecular interaction analysis on immobilized cetuximab.
- Approx. Protein Concentrations
- Total protein 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.
Erbitux® is a registered trademark of ImClone LLC, a wholly-owned subsidiary of Eli Lilly and Company.
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 Cetuximab 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 direct ELISA or as a coating antibody in PK bridging assays together with HRP-conjugated HCA221 or HCA228 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 |
|
|
| Goat anti Histidine Tag:Biotin | AHP1656B | E WB | 0.1 mg |
|
|
| Mouse anti Histidine Tag:Alk.Phos. | MCA1396A | E WB | 0.1 mg |
|
|
| Mouse anti Histidine Tag:Biotin | MCA1396B | E WB | 0.1 mg |
|
|
| Rat anti DYKDDDDK Tag:HRP | MCA4764P | E P WB | 0.1 mg |
|
|
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
|
|
| Goat anti Human IgG F(ab')2:HRP | STAR126P | E WB | 0.5 ml |
|
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Cetuximab | HCA221 | E | 0.1 mg |
|
|
| Human anti Cetuximab | HCA228 | 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 |
|
Product Specific References
Further Reading
-
Gomez, D. et al. (2013) Cetuximab therapy in the treatment of metastatic colorectal cancer: the future frontier?
Int J Surg. 11 (7): 507-13.
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