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> Anti-idiotypic Antibodies
An idiotope is the unique set of antigenic determinants (epitopes) of the variable portion of an antibody. An anti-idiotypic (Anti-ID) antibody binds to the idiotope of another antibody, usually an antibody drug, which makes it a very powerful tool for antibody drug development, especially for immunogenicity and PK/PD analysis.
To support preclinical/clinical immunogenicity and PK analysis, ACROBiosystems has developed a series of high-affinity anti-idiotypic antibodies. Our pipeline covers five hot targets including adalimumab, rituximab, cetuximab, trastuzumab, and bevacizumab. To help the drug development process, we provide assay protocols which can be applied to different application scenarios.
| Cat.No. | Antigen | Neutralizing Activity | Affinity KD, nM | Application |
|---|---|---|---|---|
| TRB-Y1b | Trastuzumab F(ab')2 | Neutralizing Antibody | 0.296 | PK bridging ELISA with TRB-Y5b Neutralizing assay Indirect ELISA |
| TRB-Y5b | Trastuzumab F(ab')2 | Neutralizing Antibody | / | PK bridging ELISA with TRB-Y1 |
| ADB-Y19 | Adalimumab F(ab’)2 | Neutralizing Antibody | 0.0013 | ADA assay Neutralizing assay Indirect ELISA |
| ADB-Y23b | Adalimumab F(ab’)2 | Non-Neutralizing Antibody | / | PK bridging ELISA with ADB-BY17 Indirect ELISA |
| ADB-BY17 | Adalimumab F(ab')2 | Neutralizing Antibody | / | PK bridging ELISA with ADB-Y23 |
| CEB-Y27 | Cetuximab F(ab’)2 | Neutralizing Antibody | 0.007 | ADA assay Neutralizing assay Indirect ELISA |
| CEB-Y28 | Cetuximab F(ab’)2 | Neutralizing Antibody | 0.0015 | ADA assay Neutralizing assay Indirect ELISA |
| CEB-Y29 | Cetuximab F(ab’)2 | Neutralizing Antibody | / | PK bridging ELISA with CEB-BY31 Neutralizing assay Indirect ELISA |
| CEB-Y31 | Cetuximab F(ab')2 | Non-Neutralizing Antibody | 0.421 | ADA assay Indirect ELISA |
| CEB-BY31 | Cetuximab F(ab')2 | Non-Neutralizing Antibody | / | PK bridging ELISA with CEB-Y29 |
| RIB-Y36 | Rituximab F(ab')2 | Neutralizing Antibody | 0.01 | ADA assay Neutralizing assay Indirect ELISA |
| RIB-Y37 | Rituximab F(ab')2 | Neutralizing Antibody | / | PK bridging ELISA with RIB-BY35 Neutralizing assay Indirect ELISA |
| BEB-Y12 | Bevacizumab F(ab’)2 | Neutralizing Antibody | 0.0828 | Neutralizing assay Indirect ELISA |
| BEB-Y9 | Bevacizumab F(ab’)2 | Neutralizing Antibody | 1.92 | ADA assay Indirect ELISA |
| BEB-Y10 | Bevacizumab F(ab')2 | Neutralizing Antibody | / | PK bridging ELISA with BEB-BY13 Neutralizing assay Indirect ELISA |
| BEB-BY13 | Bevacizumab F(ab')2 | Neutralizing Antibody | / | PK bridging ELISA with BEB-Y10 |
Therapeutic proteins such as monoclonal antibodies are currently essential in the treatment of cancer, autoimmune disease, and other diseases. Since protein has its intrinsic feature of immunogenicity owing to its structure containing potential B-cell and T-cell epitopes, therapeutic proteins have the potential to induce Anti-Drug Antibodies(ADA) even if the protein has the same amino acid sequence as endogenous human proteins. The emergence of ADA in patients can potentially lead to loss of efficacy and/or adverse events. Therefore, immunogenicity risk assessment and risk-mitigating strategies are required during the development of therapeutic protein products.
Developing a mono/multi-clonal antibody in-house as a positive control for ADA assay is extremely time-consuming. To solve this problem, ACROBiosystems developed a series of anti-drug antibody standards for ADA assays.
Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized rituximab at 1 µg/ml, added increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 10% human serum) and then added biotinylated rituximab at 2 µg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 9.7 ng/mL.
Anti-Rituximab Antibodies bridging MSD for Anti-Drug Antibody (ADA) assay development. Added the mix solution (biotinylated Rituximab at 5 µg/mL, SULFO-Rituximab at 5Nµg/mL and increasing concentrations of Anti-Rituximab Antibodies (Cat. No. RIB-Y36, 100% human serum). Detection was performed using MSD Assay with a sensitivity of 0.97 ng/mL.
Anti-Adalimumab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized adalimumab at 1 µg/ml, add increasing concentrations of Anti-Adalimumab Antibodies (Cat. No. ADB-Y19, 10% human serum) and then add biotinylated adalimumab at 5 µg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 0.6 ng/mL.
Comparison between anti-idiotypic capture ELISA and anti-idiotypic bridging ELISA for rituximab detection in patient samples. Left: anti-idiotypic capture ELISA; Right: anti-idiotypic bridging ELISA.
Detection of rituximab by bridging ELISA in serum. Immobilized Anti-Rituximab Antibodies (Cat. No. RIB-Y37) at 2 μg/ml, added increasing concentrations of Rituximab (10% human serum) and then added biotinylated Anti-Rituximab Antibodies (Cat. No. RIB-BY35) at 1 μg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 1 ng/ml.
Anti-Adalimumab Antibodies (mouse IgG1, Cat. No. ADB-Y19) captured on CM5 chip via anti-mouse antibodies surface, can bind human adalimumab with an affinity constant of 1.36 pM.
Demonstration of the specificity of Anti-Cetuximab Antibodies (Cat. No. CEB-Y28) to the cetuximab.
Reconstituted Anti-Trastuzumab Antibodies were diluted to 0.4 mg/ml, aliquoted and placed at 37°C. Aliquots were removed from 37°C at every time point and placed at 4°C along with the control. No significant loss of activity was observed.
Anti-Trastuzumab Antibodies were subjected to the indicated number of freeze-thaw cycles (FT). No significant loss of activity was observed.
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