Eculizumab antibody | AbD32311

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Human anti Eculizumab

Human Anti-Eculizumab Antibody is a recombinant, inhibitory anti-idiotypic antibody in monovalent Fab format; it is recommended as capture antibody in a PK bridging ELISA format with HCA313P for detection.

Product Type
Monoclonal Antibody
Clone
AbD32311
Isotype
HuCAL Fab monovalent
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
HCA312 E 0.1 mg
Human Anti-Eculizumab Antibody, clone AbD32311 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the free humanized monoclonal antibody eculizumab. It does not recognize free human complement C5 or eculizumab in complex with C5. The antibody can be used to measure free eculizumab or eculizumab biosimilars in patient samples.

A pair of anti-eculizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with an HRP conjugated Anti-Eculizumab Antibody in full immunoglobulin format, clone AbD32107_hIgG1 (HCA313P) as the detection antibody.

Eculizumab (Soliris) is a humanized IgG2/4 kappa antibody. The heavy chains are comprised of human IgG2 sequences in CH1, the hinge and the adjacent portion CH2, and human IgG4 sequences in the remaining part of CH2 and CH3. The drug is used to treat paroxysmal nocturnal hemoglobinuria (PNH), a rare acquired life-threatening disease of the blood characterized by destruction of red blood cells by the complement system. It also has approval as an orphan drug for treatment of atypical hemolytic uremic syndrome (aHUS). Eculizumab specifically binds to the terminal complement component 5 (C5) and inhibits the cleavage of C5 to C5a and C5b by the C5 convertase, which prevents the generation of the terminal complement complex C5b-9. Both C5a and C5b-9 cause the terminal complement-mediated events that are characteristic of PNH and aHUS. A 1:1 molar ratio of eculizumab to C5 is sufficient for C5 inhibition.

View a summary of all anti-eculizumab antibodies

Product Details

Product Form
A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in a human cell line. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH). This antibody is supplied as a liquid.
Preparation
Metal chelate affinity chromatography
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
0.01% Thiomersal
Immunogen
Eculizumab
Affinity
The monovalent intrinsic affinity of AbD32311 was measured as KD = 6 nM by real time, label free molecular interaction analysis on immobilized eculizumab
Approx. Protein Concentrations
Antibody concentration 0.5 mg/ml

Storage Information

Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
12 months from date of despatch

More Information

Acknowledgements
Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
His-tag is a registered trademark of EMD Biosciences.
Soliris is a trademark of Alexion Pharmaceuticals, Inc.
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 Eculizumab antibody

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
Clone AbD32311 can be used in an indirect ELISA or as the capture antibody for eculizumab in a bridging ELISA together with HCA313P as the detection reagent.

Protocol: PK bridging ELISA to measure free drug

Useful Reagents Available

Description Product Code Pack Size Applications List Price Quantity
Hispec Assay Diluent BUF049A 50 ml E IY
Human anti Eculizumab HCA313 0.1 mg E
Human anti Eculizumab HCA314 0.1 mg E
Human anti Eculizumab HCA315 0.1 mg E
LYNX Rapid HRP Antibody Conjugation Kit LNK002P 3 Conjugations For 400µg Antibody CJ