Anti-Alemtuzumab Antibodies

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™).

Anti-Alemtuzumab Type 1 Antibodies 

Anti-Alemtuzumab Type 1 Antibodies 

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

Anti-biotherapeutic antibody quality control and characterization


Antibodies Specific to Alemtuzumab

Product Code Clone Specificity Formats Affinity KD, nM Assay Development Recommendations
HCA174 AbD16728 Alemtuzumab Fab-V5Sx21 1.6 Indirect ELISA
PK bridging ELISA capture antibody
Inhibition assay
HCA175 AbD16942_hIgG1 Alemtuzumab Human IgG1 2.4* Indirect ELISA
PK bridging ELISA detection antibody
Inhibition assay
ADA assay
HCA175P AbD16942_hIgG1 Alemtuzumab Human IgG1
HRP labeled
  Direct ELISA
PK bridging ELISA detection antibody with HCA198
Inhibition assay
HCA198 AbD19189 Alemtuzumab Fab-FH2 0.2 PK bridging ELISA capture antibody with HCA175P
HCA199 AbD19189_hIgG1 Alemtuzumab Human IgG1 0.2* PK bridging ELISA detection antibody
Inhibition assay
ADA assay

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


Pharmacokinetic Assay - Bridging ELISA

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

Human anti-alemtuzumab antibody, product codes HCA198 and HCA175P

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.


Anti-Drug Antibody Assay - Bridging ELISA

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

Human anti-alemtuzumab antibody, product code 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)


Demonstration of Antibody Specificity


Figure 3: Specificity of anti-Alemtuzumab antibody HCA199

Human anti-alemtuzumab antibody, product code 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.