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Develop highly selective and sensitive PK and ADA assays for the humanized antibody drug alemtuzumab (MabCampath, Lemtrada) using our range of ready-made anti-idiotypic antibodies.
Table 1. Antibodies Specific to Alemtuzumab.
Specificity |
Catalog Number |
Clone |
Format |
Affinity* KD, nM |
Assay Recommendation |
Product Details |
---|---|---|---|---|---|---|
Alemtuzumab Inhibitory Type 1 |
HCA174 |
AbD16728 |
Fab-V5Sx21 |
1.6 |
PK bridging ELISA |
|
HCA175 |
AbD16942_hIgG1 |
hIgG1 HRP |
2.4 |
PK bridging ELISA |
||
HCA198 |
AbD19189 |
Fab-FH2 |
0.2 |
PK bridging ELISA |
||
HCA199 |
AbD19189_hIgG1 |
hIgG1 |
0.2 |
PK bridging ELISA |
* Affinity measured in the monovalent Fab format
1 Monovalent Fab antibody, V5- and StrepX-StrepX- tags
2 Monovalent Fab antibody DYKDDDDK- and His-6-tags
Note: The idiotope of Rat Anti-Human CD52 Antibody (MCA1642) is identical to the idiotope of Alemtuzumab
Type 1 anti-alemtuzumab antibodies inhibit the binding of the drug alemtuzumab to its target, CD52. They are ideal for development of a pharmacokinetic (PK) bridging ELISA to measure free drug. In fully human IgG1 format they are available in two affinities, and are suitable as a surrogate positive control or calibrator for an anti-drug antibody (ADA) assay.
Fig. 1. Alemtuzumab PK bridging ELISA using antibodies HCA198 and HCA175P.
Schematic image of PK bridging ELISA measuring free drug. Anti-idiotypic capture antibody, Fab format (purple), monoclonal antibody drug (gold), anti-idiotypic detection antibody, Ig format (blue), labeled with HRP.
In figure 1, Anti-Alemtuzumab Antibody (HCA198) was coated on a microtiter plate at 1.0 µg/mL and left overnight. 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 (HCA175P) at 4.0 µg/mL in HISPEC Assay Diluent (BUF049) plus QuantaBlu Fluorogenic Peroxidase Substrate.
Fig. 2. Alemtuzumab ADA bridging ELISA using antibody HCA199.
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).
In figure 2, alemtuzumab was coated at 1.5 µg/mL on a microtiter plate overnight. After washing and blocking with 5% BSA in PBST, Anti-Alemtuzumab Antibody (HCA199) was added in PBST plus 10% human serum, at the given concentrations. Detection was performed by adding HRP conjugated alemtuzumab, at 4.0 µg/mL in HISPEC Assay Diluent, 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.