Anti-Golimumab Antibodies

Antibodies for bioanalytical assays to measure golimumab and biosimilar products

Ready-made recombinant monoclonal anti-idiotypic antibodies, highly specific for the human IgG1/kappa antibody drug golimumab (Simponi®).

Type 1 anti-idiotypic antibody

Anti-Golimumab Type 1 Antibodies

The Type 1 anti-golimumab antibodies inhibit the binding of the drug golimumab to its target, human 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: HCA240 and HCA241

Type 2 Anti-Golimumab Antibodies

Anti-Golimumab Type 2 Antibodies

The Type 2 anti-golimumab antibody does not inhibit the binding of golimumab to TNFα and can therefore detect total drug – free, partially bound and fully bound. These fully human antibodies can be used to measure the golimumab levels in patient sera.
Product codes: HCA242 and HCA243

 

Type 3 Anti-Golimumab/TNF-a Antibodies

Anti-Golimumab Drug/Target Complex Type 3 Antibodies

The Type 3 antibodies specifically recognize the golimumab/TNFα complex. They detect golimumab only when it is bound to TNFα. These antibodies can be used to develop PK assays that avoid the bridging format and allow detection of golimumab bound to its target in serum as opposed to free golimumab.
Product codes: HCA244 and HCA245


The recombinant anti-golimumab antibodies are generated using the HuCAL® antibody library and a proprietary improved form of phage display. Antibody generation using HuCAL technology results in highly specific and sensitive antibodies that are ideal for PK and PD assays. Furthermore, antibodies are fully human and so are also suitable 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 Golimumab

Product
Code
Clone

Antibody
Specificity

Binding Type
Format
Affinity KD, nM
Assay Development 
Recommendations
AbD20710
Golimumab
Type 1
Fab-FH1
2.3
Direct and Indirect ELISA 
PK bridging ELISA with HCA241
AbD20692_hIgG1
Golimumab
Type 1
Human
IgG1
2.82
Direct and Indirect ELISA
PK bridging ELISA with HCA240 
ADA bridging ELISA
AbD20897
Golimumab
Type 2
Fab-FH1
7
PK bridging ELISA, detection antibody
AbD20897_hIgG1
Golimumab
Type 2
Human
IgG1
72
PK bridging ELISA, detection antibody
AbD20893
Golimumab/TNFα
Type 3
Fab-FH1
53
(on complex)
PK ELISA antigen capture format
AbD20893_hIgG1
Golimumab/TNFα
Type 3
Human
IgG1
532
(on complex)
PK ELISA antigen capture format

Table 1 Antibody Specifications
F=DYKDDDDK-tag H=His-6-tag  
Affinity measured in the monovalent Fab format

Related Products

Golimumab drug target Recombinant human TNF alpha (PHP051)
Golimumab 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

PK assay, bridging format, using antibodies HCA240 and HCA241

Figure 1: Golimumab PK ELISA bridging format using antibodies HCA240 and HCA241

Human anti-golimumab antibodies, product codes HCA240 and HCA241

Figure 1: Anti-golimumab antibody, clone AbD20710 (HCA240) 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. Golimumab titrated into 10% human serum was added. Detection was performed using HRP conjugated anti-golimumab antibody clone AbD20692_hIgG1* (HCA241) 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 golimumab HCA240 and HCA241


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

PK assay in antigen capture format ELISA, using Type 3 antibody

Figure 2: Golimumab PK ELISA antigen capture format using antibody HCA244

Human anti-golimumab antibody, product code HCA244

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, golimumab spiked into 10% human serum was added in the given concentrations. Detection was performed by using anti-golimumab/TNFα complex specific antibody, clone AbD20893* (HCA244) at a concentration of 2 µg/ml, followed by anti-Penta Histidine Tag:HRP (MCA5995P) in HISPEC assay diluent (BUF049) and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

Protocol: PK antigen capture ELISA protocol golimumab


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 assay development, using Type 1 anti-golimumab antibody HCA241

Figure 3: ADA bridging format using type 1 anti-golimumab antibody HCA241

Human anti-golimumab antibody, product code HCA241

Figure 3: Golimumab was coated at 1 µg/ml on a microtiter plate overnight. After washing and blocking with PBST+5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of anti-golimumab antibody, clone AbD20692_hIgG1 (HCA241). Detection was performed using HRP conjugated golimumab*. Data are shown as the mean of three measurements.  


Inhibition ELISA
Inhibition of golimumab binding to TNFα

Inhibition of golimumab binding to TNF alpha 

Schematic image of inhibition assay. Drug target capture antigen (red), monoclonal antibody drug (gold), anti-IgG Fc specific detection antibody (gray) labeled with HRP

Inhibition of golimumab to TNF-a by Type 1 antibody HCA240, but not by Type 2 antibody HCA242

Figure 4: Inhibition of golimumab binding to TNFα by type 1 antibody HCA240, but not by type 2 antibody HCA242

Human anti-golimumab antibodies, product codes HCA240 and HCA242

Figure 4: A microtiter plate was coated over night with recombinant human TNFα (PHP051) at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of golimumab (0.3 µg/ml) plus increasing concentrations of either anti-golimumab type 1 antibody, clone AbD20710 (HCA240) or anti-golimumab type 2 antibody, clone AbD20897 (HCA242) was added. Free golimumab, still capable of binding to the human TNFα coated plate, was detected using HRP conjugated mouse anti human IgG (Fc) CH2 domain (MCA647P) and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements. 


Demonstration of Antibody Specificity 

Specificity of Type 3 antibody HCA245 to drug-target complex

Figure 5: Specificity of drug-target complex antibody HCA245

Human anti-golimumab antibody, product code HCA245

Figure 5: Demonstration of the specificity of antibodies to the golimumab/TNFα complex. Type 3 antibodies (HCA244 and HCA245) will detect golimumab only when bound to its target TNFα (drug-target complex binders). These antibodies do not detect either golimumab or TNFα when they are not in a drug-target complex. Various antigens were coated at 5.0 µg/ml on a microtiter plate overnight. After washing and blocking with PBST+5% BSA, detection was performed using HRP conjugated anti-golimumab (drug/target complex) antibody, clone AbD20893_hIgG1* (HCA245) at a concentration of 2 μg/ml and QuantaBlu fluorogenic peroxidase substrate.

*Antibodies were conjugated to HRP using a LYNX Rapid HRP Conjugation Kit® (LNK001P-LNK006P).


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.

Simponi® is a registered trademark of Centocor Ortho Biotech Inc. HuCAL® is a registered trademark of MorphoSys AG. QuantaBlu™ is a trademark of Thermo Fisher Scientific.