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Anti-idiotypic Antibodies

Anti-idiotypic (Anti-ID) antibodies recognize unique epitopes in the variable region of a therapeutic antibody. In pharmacokinetic (PK) assays, they can be used as capture or detection reagents for drug-specific quantification in biological samples. For anti-drug antibody (ADA) assays, drug-specific anti-ID antibodies can provide positive controls for immunogenicity assay development and evaluation.

ACROBiosystems offers anti-idiotypic antibodies for a range of therapeutic antibodies, including adalimumab, rituximab, cetuximab, trastuzumab, and bevacizumab. Application data and assay protocols are available to support product selection and method development.

Anti-idiotypic Antibody Products List

Cat. No.TargetDrug NameProduct DescriptionOrder Now
TNF-α
TNF-α
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TNF-α
TNF-α
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TNF-α
TNF-α
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BCMA
BCMA
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BCMA
BCMA
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BCMA
BCMA
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BCMA
BCMA
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VEGF-A
VEGF-A
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VEGF-A
VEGF-A
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VEGF-A
VEGF-A
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VEGF-A
VEGF-A
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EGF R
EGF R
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EGF R
EGF R
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EGF R
EGF R
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EGF R
EGF R
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EGF R
EGF R
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TROP-2
TROP-2
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TROP-2
TROP-2
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TROP-2
TROP-2
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TROP-2
TROP-2
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Her2
Her2
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Her2
Her2
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Her2
Her2
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Nectin-4
Nectin-4
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Nectin-4
Nectin-4
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Her2
Her2
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Her2
Her2
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Her2
Her2
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Siglec-2
Siglec-2
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Siglec-2
Siglec-2
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Siglec-2
Siglec-2
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CD79B
CD79B
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CD79B
CD79B
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CD79B
CD79B
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CD20
CD20
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CD20
CD20
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CD20
CD20
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TROP-2
TROP-2
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TROP-2
TROP-2
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TROP-2
TROP-2
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Her2
Her2
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Her2
Her2
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Her2
Her2
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Two Key Applications

  • Application 1: Drug-Specific Quantification for PK Analysis
  • Application 2: Drug-Specific Positive Controls for ADA Assays

Application 1: Drug-Specific Quantification for PK Analysis

Assays that rely on generic anti-human IgG reagents may show elevated background in complex biological matrices because of interference from endogenous immunoglobulins. By recognizing the variable region of a specific therapeutic antibody, anti-idiotypic antibodies help distinguish the drug from endogenous IgG and can be used as capture reagents, detection reagents, or matched pairs.
Different analytes call for different types of anti-idiotypic antibodies:
Free drug: Antigen-blocking anti-idiotypic antibodies are used to detect unbound therapeutic antibody.
Total antibody drug: Antigen non-blocking anti-idiotypic antibodies can detect the therapeutic antibody across free and target-bound forms.
Target-bound drug: Antibodies that specifically recognize the therapeutic antibody–target complex are used to detect the bound form.
Rituximab PK Assay Case

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.

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Comparison of 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.

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.

Application 2: Drug-Specific Positive Controls for ADA Assays

ADA assays typically require a positive control tailored to the therapeutic being evaluated. Anti-idiotypic antibodies developed against that drug can serve as positive controls in bridging ELISA and MSD assays. Antibodies with confirmed neutralizing activity may also be used in neutralizing antibody assays
Anti-Rituximab Antibody in ADA Assays
The anti-rituximab antibody shown below was tested in both bridging ELISA and MSD formats. The bridging ELISA achieved a sensitivity of 9.7 ng/mL in 10% human serum, while the bridging MSD assay achieved a sensitivity of 0.97 ng/mL in 100% human serum.

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.

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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.

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Anti-Adalimumab Antibody in ADA Assays

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.

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Validation Data

Binding Affinity
SPR confirms binding between the anti-idiotypic antibody and its corresponding therapeutic antibody, providing affinity data for product selection and assay development.

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.

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Specificity Data
Cross-reactivity is assessed using the therapeutic antibody, its target, different IgG subclasses, and other antibodies of the same subclass to confirm drug-specific recognition.

Demonstration of the specificity of Anti-Cetuximab Antibodies (Cat. No. CEB-Y28) to the cetuximab.
 

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Stability Data
Accelerated and freeze-thaw studies are used to evaluate the stability of representative products.

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 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.

Anti-Idiotypic Antibody Development Service

For projects not covered by off-the-shelf products, ACROBiosystems provides custom services spanning antigen preparation, monoclonal or polyclonal anti-idiotypic antibody development, and PK or immunogenicity assay kit development. These services can support programs involving monoclonal antibodies, bispecific antibodies, antibody-drug conjugates, CAR-T therapies, and peptides.
Depending on the assay objective, custom programs can include antigen-blocking antibodies, antigen non-blocking antibodies, therapeutic antibody–target complex-specific antibodies, and matched antibody pairs. Polyclonal anti-idiotypic antibodies can also be developed as positive controls for ADA assays. Each program can be tailored to the therapeutic format, target analyte, and assay platform.

Learn About Anti-Idiotypic Antibody Development Services

  • Background
  • Anti-idiotypic Antibody Products List
  • Two Key Applications
  • Validation Data
  • Anti-Idiotypic Antibody Development Service
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