Anti-Dupilumab Antibodies

Anti-Dupilumab Inhibitory Antibodies (Type 1)

Type 1 anti-dupilumab antibodies inhibit the binding of the drug dupilumab to its target, interleukin-4 subunit α (IL-4Rα) receptor. 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.

TrailBlazer Antibodies, clones TZA014 AbD47091ad, TZA015 AbD47092ad have a SpyTag2 incorporated at the C-terminus of the antibody heavy chain. In a fast reaction that simply requires mixing at room temperature, these antibodies can form a covalent isopeptide bond to a SpyCatcher reagent, enabling site-directed conjugation or fast conversion to a bivalent Fab or a full-length Ig-like format within an hour.

Table 1. Antibodies Specific to Dupilumab.

Specificity
Binding Type

Catalog Number

Clone

Format

Affinity* KD, nM

Assay Recommendation

Product Details

Dupilumab Inhibitory Type 1

HCA383

AbD47090ia

Human IgG1

0.4

ADA assay

Order HCA383

HCA384

AbD47092ia

Human IgG1

0.3

ADA assay

Order HCA384

TZA014

AbD47091ad

Fab-F-Spy2-H

0.8

PK bridging ELISA

Order TZA014

TZA015

AbD47092ad

Fab-F-Spy2-H

0.3

PK bridging ELISA

Order TZA015

ELISA Protocols to Get You Started

PK Bridging ELISA


Fig. 1. A microtiter plate was coated over night with human anti dupilumab (clone AbD47091ad, TZA014) at a concentration of 1 µg/ml.

Fig. 1. A microtiter plate was coated over night with human anti-dupilumab (clone AbD47091ad, TZA014) at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, 10% human serum was added spiked with increasing concentrations of dupilumab. Detection was performed using HRP-conjugated human anti-dupilumab (clone AbD47092pap, TZA015P) at a concentration of 0.5 µg/mL in HISPEC assay diluent and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

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, Fab format (purple), labeled with HRP.


Anti-Drug Antibody Bridging ELISA


Fig. 2. A microtiter plate was coated over night with dupilumab at a concentration of 1 µg/ml.

Fig. 2. A microtiter plate was coated over night with dupilumab at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of human anti-dupilumab clone AbD47090ia (HCA383) and AbD47092ia (HCA384). Detection was performed using HRP-conjugated dupilumab at a concentration of 2 µg/ml in HISPEC assay diluent and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

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).


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