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Develop highly selective and sensitive PK and ADA assays for the monoclonal antibody drug eculizumab (Soliris) using ready-made anti-idiotypic antibodies.
Table 1. Antibodies Specific to Eculizumab, Human C5 and Eculizumab–Human C5 complex
Specificity |
Catalog Number |
Clone |
Format |
Affinity* KD, nM |
Assay Recommendation |
Product Details |
---|---|---|---|---|---|---|
Eculizumab Inhibitory Type 1 |
HCA312 |
AbD32311 |
Fab-FH1 |
6 |
PK bridging ELISA |
|
HCA313 |
AbD32107_hIgG1 |
Human IgG1 HRP |
0.6 |
PK bridging ELISA ADA control |
||
HCA314 |
AbD32311_hIgG1 |
Human IgG1 |
6 |
ADA control |
||
HCA315 |
AbD32109_hIgG1 |
Human IgG1 |
3 |
ADA control |
||
Eculizumab/ Human C5 Complex Specific Type 3 |
HCA339 |
AbD32334 |
Fab-FH1 |
57 |
PK bridging ELISA (capture) |
|
Anti-Human C5 Antibody |
TZA034
|
AbD36700ad |
Fab-F-Spy2-H |
Not determined |
Detection antibody for Type 3 assay |
|
TZA034P |
AbD36700pap |
Fab2-FH-X22-HRP |
Not determined |
Detection antibody for Type 3 assay |
* Affinity measured in the monovalent Fab format
1 Fab-F-Spy2-H = Monovalent Fab antibody with (F) DYKDDDDK, (Spy2) SpyTag version 2, and (H) His-6-tags
2 Fab2-FH-X22-HRP = Bivalent Fab antibody with (F) DYKDDDDK, (H) His-6-tags and Fab-SpyTag coupled to HRP conjugated BiCatcher
Type 1 anti-eculizumab antibodies inhibit the binding of the drug eculizumab to its target, human complement C5 protein (C5). They are ideal for development of a pharmacokinetic (PK) bridging ELISA to measure free drug. In fully human IgG1 format they are suitable as a surrogate positive control or calibrator for an anti-drug antibody (ADA) assay.
The Type 3 antibody specifically recognizes the drug-target complex, detecting eculizumab only when it is bound to human C5. It is suitable as a capture antibody in a PK bridging ELISA in combination with anti-C5 antibodies for detection.
Fig. 1. Eculizumab PK bridging ELISA using antibodies HCA312 and HCA313P.
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.
Fig. 2. Eculizumab PK bridging ELISA using HCA399 and TZA034P.
Fig. 3. Eculizumab ADA bridging ELISA using antibody HCA313, HCA314, or HCA315.
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).
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.