Anti-Adalimumab Antibodies

Ideal for bioanalytical assays supporting preclinical research and clinical drug monitoring

Ready-made recombinant, anti-idiotypic monoclonal antibodies, highly specific for the human antibody drug adalimumab (HUMIRA®).

Type 1 anti-idiotypic antibody

Anti-Adalimumab Type 1 Antibodies

The Type 1 anti-adalimumab antibodies inhibit the binding of the drug adalimumab to its target, TNFα, and therefore detect free drug. These antibodies are ideal for use in direct or indirect 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: HCA202, HCA203, HCA204, HCA204P, HCA205
 

Type 3 anti-drug-target complex specific antibody

Anti-Adalimumab Drug/Target Complex Type 3 Antibodies

The Type 3 antibodies specifically recognize the adalimumab/TNFα complex. These antibodies detect adalimumab only when it is bound to TNFα.  They do not bind to free adalimumab or TNFα. With these antibodies, PK assays can be built that avoid the bridging format. In addition, they allow detection of adalimumab bound to its target in serum as opposed to free adalimumab.
Product codes: HCA206, HCA207, HCA231, HCA232, HCA232P

The recombinant anti-adalimumab antibodies are generated using the HuCAL® antibody library and a modified and improved form of phage display. Antibody generation using HuCAL technology results in highly specific and sensitive antibodies, which are ideal for PK and PD assays. Antibodies are fully human, so are also perfect as controls or calibrators for ADA assays. The in vitro production of recombinant antibodies means that there is a consistent, secure supply throughout preclinical development and clinical trials, avoiding the need to develop new reagents.

More information about anti-idiotypic antibodies and their binding types and properties

Anti-biotherapeutic antibody quality control and characterization


Antibodies Specific to Adalimumab

Product Code Clone Antibody Specificity Binding Type Format Affinity KD, nM Assay Development Recommendations
HCA202 AbD18654 Adalimumab Type 1 Fab-FH1 0.15 Direct and indirect ELISA
PK bridging ELISA with HCA204
Inhibition ELISA
HCA203 AbD18654_hIgG1 Adalimumab Type 1 Human IgG1 0.15* Direct ELISA
ADA assay
Inhibition ELISA
HCA204 AbD18655_hIgG1 Adalimumab Type 1 Human IgG1 0.06* PK bridging ELISA with HCA202
ADA assay
Inhibition ELISA
HCA204P AbD18655_hIgG1 Adalimumab Type 1 Human IgG1
HRP labeled
0.06* PK bridging ELISA with HCA202
Inhibition ELISA
HCA205 AbD18664_hIgG1 Adalimumab Type 1 Human IgG1 10.5* Direct ELISA
ADA assay
HCA206 AbD18754 Adalimumab/TNFα Type 3 Fab-FH1 67 PK antigen capture format
HCA207 AbD18754_hIgG1 Adalimumab/TNFα Type 3 Human IgG1 67* PK antigen capture format
HCA2312 AbD20350 Adalimumab/TNFα Type 3 Fab-FH1 3.2 PK antigen capture format
HCA2322 AbD20349_hIgG1 Adalimumab/TNFα Type 3 Human IgG1 6.7* PK antigen capture format
HCA232P2 AbD20349_hIgG1 Adalimumab/TNFα Type 3 Human IgG1
HRP labeled
6.7* PK antigen capture format

Table 1 Antibody Specifications

1F=DYKDDDDK-tag H=His-6-tag  
2HCA231 and HCA232/HCA232P are affinity matured versions of clone 18754 (HCA206 and HCA207, respectively)
*Affinity measured in the monovalent Fab form

Related Products

Adalimumab drug target Recombinant human TNF alpha (PHP051)
Adalimumab drug isotype control Recombinant human IgG1 kappa (HCA192)


Pharmacokinetic Assay - Bridging Format

Schematic image of anti-idiotypic antibodies in PK bridging ELISA format

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

Adalimumab PK bridging ELISA using antibodies HCA202 and HCA204/HCA204P

Figure 1: Adalimumab PK ELISA bridging format using antibodies HCA202 and HCA204P

Human anti-adalimumab antibodies, product codes HCA202 and HCA204

Figure 1: Anti-adalimumab antibody, clone AbD18654 (HCA202) was coated on a microtiter plate at 1.0 µg/ml and left over night. Washing and blocking was performed with 5% BSA in PBST. Adalimumab titrated into 10% human serum was added. Detection was performed using HRP conjugated anti-adalimumab antibody clone AbD18655_hIgG1 (HCA204P) diluted to 2 µg/ml in HISPEC assay diluent (BUF049), plus QuantaBlu™ fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

Protocol: PK bridging ELISA protocol adalimumab HCA202 and HCA204P


Pharmacokinetic Assay – Antigen Capture Format

Schematic image of anti-drug/target complex specific antibody in PK ELISA antigen capture format

Schematic image of PK antigen capture format ELISA. Drug target (red), monoclonal antibody drug (gold), drug target complex detection antibody, Fab format (purple), labeled with HRP

Adalimumab PK ELISA antigen capture format using antibody HCA231

Figure 2: Adalimumab PK ELISA antigen capture format using antibody HCA231

Human anti-adalimumab antibody, product code HCA231

Figure 2: Recombinant human TNFα (PHP051) was coated on a microtiter plate at 5 µg/ml. After washing and blocking with 5% BSA in PBST, adalimumab spiked into 10% human serum was added in the given concentrations. After washing, either unlabeled or HRP labeled anti-adalimumab/TNFα complex specific antibody, clone AbD20350 (HCA231) was added at 2µg/ml in PBST. Unlabeled HCA231 was detected by adding anti-Penta Histidine Tag:HRP (MCA5995P) in HISPEC assay diluent (BUF049) and QuantaBlu fluorogenic peroxidase substrate. HRP-labeled HCA231 was detected directly with QuantaBlu substrate. Data are shown as the mean of three measurements. Antibodies were conjugated to HRP using a LYNX Rapid Conjugation Kit® (LNK001P-LNK006P).

Protocol: PK antigen capture ELISA protocol adalimumab


Anti-Drug Antibody Assay - Bridging ELISA

Schematic image of fully human anti-idiotypic antibody in anti-drug antibody (ADA) assay

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)

ADA Bridging ELISA using anti adalimumab antibodies

Figure 3: ADA assay, bridging format using antibodies HCA203, HCA204 and HCA205

Human anti-adalimumab antibodies, product codes HCA203, HCA204 and HCA205

Figure 3: Adalimumab was coated at 1 µg/ml on a microtiter plate overnight. After washing and blocking with 5% BSA in PBST, the anti-adalimumab antibodies in human IgG1 format (black squares adalimumab antibody clone AbD18654_hIgG1 (HCA203); red circles adalimumab antibody clone AbD18655_hIgG1 (HCA204); green triangles adalimumab antibody clone AbD18664_hIgG1 (HCA205) spiked in 10% human serum in the given concentrations were added. Detection was performed by adding HRP conjugated adalimumab, 2 µg/ml in HISPEC assay diluent (BUF042), plus QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

Protocol: ADA bridging ELISA protocol adalimumab


Inhibition ELISA

Inhibition of adalimumab binding to TNFa using antibodies HCA203 and HCA207

Figure 4: Inhibition of adalimumab binding to TNFα using antibodies HCA203 and HCA207

Human anti-adalimumab antibodies, product codes HCA203 and HCA207

Figure 4: Recombinant human TNFα (PHP051) was coated on a microtiter plate at 1.0 µg/ml, followed by a pre-incubated mixture of the anti-adalimumab antibody, clone AbD18654_hIgG1 (HCA203) titrated into a constant amount of adalimumab (0.3 µg/ml). Free adalimumab, still capable of binding to the plate, was detected using HRP conjugated anti-adalimumab/TNFα complex antibody, clone AbD18754_hIgG1 (HCA207). Data are shown as the mean of three measurements.


Determination of Antibody Specificity

Demonstration of the specificity of antibody HCA232 to the adalimumab/TNFa complex

Figure 5: Demonstration of the specificity of antibody HCA232 to the adalimumab/TNFα complex. 

Human anti-adalimumab antibody, product code HCA232P

Figure 5: Type 3 antibodies (HCA206, HCA207, HCA231 and HCA232) will detect adalimumab only when bound to its target TNFα. A microtiter plate was coated over night with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP-conjugated anti-adalimumab/TNFα complex-specific antibody, clone AbD20349_hIgG1 (HCA232P) and 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. 

HUMIRA® is a registered trademark of Abbott Laboratories. HuCAL® is a registered trademark of MorphoSys AG. QuantaBlu™ is a trademark of Thermo Fisher Scientific.