Antibodies for bioanalysis and drug monitoring of alemtuzumab

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.

  • Inhibitory antibodies specific to alemtuzumab
  • Fully human surrogate positive control or calibrator
  • Sequence defined, well characterized reagents with secure supply
  • Recombinant production, stringent quality control for batch to batch consistency
  • Non-animal-derived antibodies, reducing the use of animals in science

Table 1. Antibodies Specific to Alemtuzumab.

Specificity
Binding Type

Catalog Number

Clone

Format

Affinity* KD, nM

Assay Recommendation

Product Details

Alemtuzumab Inhibitory

Type 1

HCA174

AbD16728

Fab-V5Sx21

1.6

PK bridging ELISA

Order HCA174

HCA175
HCA175P

AbD16942_hIgG1

hIgG1

HRP

2.4

PK bridging ELISA
ADA control

Order HCA175
Order HCA175P

HCA198

AbD19189

Fab-FH2

0.2

PK bridging ELISA

Order HCA198

HCA199

AbD19189_hIgG1

hIgG1

0.2

PK bridging ELISA
ADA control

Order HCA199

* 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


Anti-Alemtuzumab Inhibitory Antibodies (Type 1)

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.


PK Bridging ELISA

Fig. 1. Alemtuzumab PK bridging ELISA using antibodies HCA198 and HCA175P.

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.

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.


ADA Bridging ELISA

Fig. 2. Alemtuzumab ADA bridging ELISA using antibody HCA199.

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).

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.


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