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 Inhibitory Antibodies

These inhibitory (Type 1) antibodies prevent the binding of the drug golimumab to its target, human TNFα, and therefore detect free drug. They 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.
Catalog Numbers: HCA240, HCA241, HCA286, HCA287, HCA287P.

Type 2 Anti-Golimumab Antibodies

Anti-Golimumab Non-Inhibitory Antibodies

These non-inhibitory (Type 2) antibodies do 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.
Catalog Numbers: HCA242, HCA243, HCA289, HCA290, HCA290P.

 

Type 3 Anti-Golimumab/TNF-a Antibodies

Anti-Golimumab Drug/Target Complex Specific Antibodies

These antibodies (Type 3) specifically recognize the golimumab/TNFα complex. They detect golimumab only when it is bound to TNFα. They can be used to develop PK assays that avoid the bridging format and allow detection of golimumab bound to its target, as opposed to free golimumab. 
Catalog Numbers: HCA245, HCA274.


The recombinant anti-golimumab antibodies are generated using the HuCAL® antibody library and CysDisplay®, a proprietary improved form of phage display. Antibody generation using HuCAL technology results in highly specific and sensitive antibodies that are ideal for PK 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

Catalog Number
Clone

Antibody
Specificity

Binding Type
Format
Affinity KD, nM
Assay Development 
Recommendations
AbD20710
Golimumab
Type 1
Fab-FH1
2.3
PK bridging ELISA (capture)
AbD20692_hIgG1
Golimumab
Type 1
Human
IgG1
2.82
PK bridging ELISA (detection)
ADA bridging ELISA
HCA286 AbD254293 Golimumab Type 1 Fab-FH1 1.0
PK bridging ELISA (capture) with HCA287P
HCA287 AbD25418_hIgG13 Golimumab Type 1 Human
IgG1
0.12
PK bridging ELISA (detection) with HCA286
ADA bridging ELISA
HCA287P AbD25418_hIgG13 Golimumab Type 1 Human
IgG1
HRP
labeled
0.12 PK bridging ELISA (detection) with HCA286
AbD20897
Golimumab &
Golimumab/TNFα complex
Type 2
Fab-FH1
7
PK bridging ELISA (capture)
AbD20897_hIgG1
Golimumab &
Golimumab/TNFα complex
Type 2
Human
IgG1
72
PK bridging ELISA (detection)
HCA289 AbD254553 Golimumab &
Golimumab/TNFα complex
Type 2 Fab-FH1 0.9 PK bridging ELISA (capture) with HCA290P
HCA290 AbD25451_hIgG13 Golimumab &
Golimumab/TNFα complex
Type 2 Human IgG1 0.42 PK bridging ELISA (detection) with HCA289
HCA290P AbD25451_hIgG13 Golimumab &
Golimumab/TNFα complex
Type 2 Human IgG1
HRP labeled
0.42 PK bridging ELISA (detection) with HCA289
HCA274 AbD257053 Golimumab/TNFα 
complex
Type 3 Fab-FH1 6
(on complex)
PK ELISA antigen capture format
AbD20893_hIgG1
Golimumab/TNFα
complex
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
Affinity matured antibody: HCA286 is derived from AbD20710 (HCA240); HCA287 is derived from AbD20692 (HCA241); HCA289 and HCA290 are derived from AbD20897 (HCA242); HCA274 is derived from AbD20893 (HCA245)

Related Products

Golimumab drug target Recombinant Human TNF Alpha (PHP051)
Golimumab drug isotype control Recombinant Human IgG1 Kappa (HCA192)


Pharmacokinetic Assay - Bridging ELISA

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.

Fig. 1. Golimumab PK bridging ELISA bridging using antibodies HCA286 and HCA287P

Fig. 1. Golimumab PK bridging ELISA using antibodies HCA286 and HCA287P.

Measurement of Free Drug

In Figure 1, a microtiter plate was coated overnight with Human Anti-Golimumab Antibody clone AbD25429 (HCA286) 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 golimumab. Detection was performed using HRP conjugated Anti-Golimumab Antibody clone AbD25418_hIgG1 (HCA287P) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.

Protocol: PK bridging ELISA protocol golimumab HCA286 and HCA287P


Schematic image of PK Bridging ELISA with type 2 antibodies

Schematic image of PK Bridging ELISA with type 2 antibodies. Anti-idiotypic capture antibody, Fab format (purple), monoclonal antibody drug (gold), anti-idiotypic detection antibody, Ig format (blue), labeled with HRP. Assay measures total drug (free, partially or fully bound).

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

Fig. 2. Golimumab PK bridging ELISA using antibodies HCA289 and HCA290P.

Measurement of Total Drug

In Figure 2, a microtiter plate was coated overnight with Anti-Golimumab Antibody clone AbD25455 (HCA289) 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 golimumab. Detection was performed using HRP conjugated Anti-Golimumab Antibody clone AbD25451_hIgG1 (HCA290P) 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.

Protocol: PK bridging ELISA protocol golimumab HCA289 and HCA290P


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

Fig. 3. Golimumab PK ELISA antigen capture format using antibody HCA274.

Measurement of Bound Drug

In Figure 3, Recombinant Human TNFα (PHP051) was coated on a microtiter plate at 5 µg/ml. After washing and blocking with PBST + 5% BSA, 10% human serum was added spiked with increasing concentrations of golimumab. Detection was performed by using Anti-Golimumab/TNFα Complex Specific Antibody, clone AbD25705 (HCA274) at a concentration of 2 µg/ml, followed by Mouse Anti-Penta Histidine Tag:HRP Antibody (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

Fig. 4. ADA bridging ELISA using type 1 Anti-Golimumab Antibody HCA287.


In Figure 4, a microtiter plate was coated overnight with golimumab 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 Anti-Golimumab Antibody clone AbD25418_hIgG1 (HCA287). Detection was performed using HRP conjugated golimumab and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.

Protocol: ADA bridging ELISA protocol golimumab HCA287


Inhibition ELISA

Inhibition of Golimumab Binding to TNFα

Fig. 5. Inhibition of golimumab binding to TNFα by type 1 antibody HCA287, but not by type 2 antibody HCA289.


Inhibition of Golimumab Binding to TNFα

In Figure 5, a microtiter plate was coated overnight 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 Anti-Golimumab Antibody, clone AbD25418 or clone AbD25455 was added (both antibodies in monovalent Fab format in this assay). Free golimumab, still capable of binding to the TNFα coated plate was detected using Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.


Demonstration of Antibody Specificity 

Drug/target complex specific (Type 3) antibodies HCA274 and HCA245 only detect golimumab when bound to its target TNFα. These antibodies do not detect either golimumab or TNFα when they are not in a drug/target complex.

Drug/target complex specific (Type 3) antibodies HCA274 and HCA245

Fig. 6. Specificity of drug/target complex antibody HCA274.

In Figure 6, a microtiter plate was coated overnight with various antigens at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Anti-Golimumab Antibody clone AbD25705 (HCA274) at a concentration of 2 µg/ml followed by Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent 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.