Antibodies for bioanalytical assays to measure alemtuzumab and biosimilar products
Ready-made recombinant monoclonal anti-idiotypic antibodies, highly specific for the humanized antibody drug alemtuzumab (MabCampath®, LEMTRADA™).
The Type 1 anti-Alemtuzumab antibodies inhibit the binding of the drug alemtuzumab to its target, CD52, and therefore detect free drug. These antibodies are ideal for use in direct ELISA, pharmacokinetic (PK) bridging assays and to develop and calibrate immune response (IR) assays to measure the anti-drug antibody (ADA) response in patient sera.
Product codes: HCA174, HCA175, HCA175P, HCA198, HCA199
These recombinant, monoclonal, anti-idiotypic antibodies are generated using the HuCAL® antibody library and a proprietary method of phage display, which results in highly specific and sensitive reagents, ideal for PK and PD assays. Because these antibodies are fully human, they are also perfect as a refernce standard for ADA assays when in full immunoglobulin format. The in vitro production of recombinant antibodies results in a consistent, secure supply of these critical reagents throughout preclinical development and clinical trials.
More information about anti-idiotypic antibodies and their binding types and properties
|Product Code||Clone||Specificity||Formats||Affinity KD, nM||Assay Development Recommendations|
PK bridging ELISA capture antibody
PK bridging ELISA detection antibody
PK bridging ELISA detection antibody with HCA198
|HCA198||AbD19189||Alemtuzumab||Fab-FH2||0.2||PK bridging ELISA capture antibody with HCA175P|
PK bridging ELISA detection antibody
Table 1 Antibody Specifications
1 Monovalent Fab antibody, V5-tag and StrepX-StrepX-tag
2 Monovalent Fab antibody, DYKDDDDK-tag and His-6-tag
* Affinity measured in the monovalent Fab form
Note: The idiotope of rat anti-CD52 (MCA1642) is identical to Alemtuzumab
Schematic image of PK Bridging ELISA. Anti-idiotypic capture antibody, Fab format (purple), monoclonal antibody drug (gold), anti-idiotypic detection antibody, Ig format (blue), labeled with HRP
Figure 1: PK bridging ELISA using, antibodies HCA198 and HCA175
Figure 1: Anti-alemtuzumab antibody clone AbD19189 (HCA198) was coated on a microtiter plate at 1.0 µg/mL and left over night. After washing and blocking with 5% BSA in PBST, alemtuzumab was spiked in 10% human serum and added in increasing concentrations as indicated. Detection was performed using HRP conjugated anti-alemtuzumab antibody, clone AbD16942_hIgG1 (HCA175P), 4.0 µg/mL in HISPEC buffer, plus QuantaBlu™ fluorogenic peroxidase substrate.
Schematic image of ADA bridging assay. Monoclonal antibody drug as capture antibody and detection antibody labeled with HRP (gold), fully human anti-idiotypic antibody, Ig format (blue)
Figure 2: ADA bridging ELISA using antibody HCA199
Figure 2: Alemtuzumab was coated at 1.5 µg/mL on a microtiter plate over night. After washing and blocking with 5% BSA in PBST, anti-alemtuzumab antibody, clone AbD19189_hIgG1 (HCA199) was added in PBST plus 10% human serum, at the given concentrations. Detection was performed adding HRP conjugated alemtuzumab*, 4.0 µg/mL in HISPEC buffer, plus QuantaBlu fluorogenic peroxidase substrate.
*Antibody conjugated to HRP using a LYNX Rapid Conjugation Kit® (product codes LNK001P - LNK006P)
Figure 3: Specificity of anti-Alemtuzumab antibody HCA199
Figure 3: Demonstration of the specificity of antibodies to alemtuzumab. Antigens were coated at 5.0 µg/mL on a microtiter plate over night. After washing and blocking with 5% BSA, anti-alemtuzumab antibody HCA199, 20 µL from a 2.0 µg/mL solution in HISPEC buffer, was added. Detection was performed using HRP conjugated anti-Penta Histidine tag antibody (MCA5995P) in HISPEC buffer, plus QuantaBlu fluorogenic peroxidase substrate.
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.
LEMTRADA™ and Campath® are trademarks of Genzyme Corporation. HuCAL® is registered trademark of MorphoSys AG. QuantaBlu™ is a trademark of Thermo Fisher.