Conjugated Antibody Determination
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Conjugated antibody refers to antibody forms with at least one conjugated payload. Though the correlation between levels of conjugated antibody and efficacy or toxicity of an ADC has not yet been fully established, considerable interests have been directed to measurement of the conjugated antibody, which is recognized as one of the important active species of ADCs. Creative Proteomics utilizes our advanced LBA and LC-MS/MS technology platforms to accurately quantify conjugated antibodies to ensure that our clients are provided with complete pharmacokinetic and immunogenicity data to support their antibody drug conjugates-related research processes.
The Role and Significance of Conjugated Antibody Quantification
After ADC enters the body, the cytotoxic effector molecules can be gradually dissociated from the main structure by enzymatic or chemical reactions to obtain total antibody (Tab), conjugated antibody (ADC) and free effector molecules (unconjugated drug). Among them, conjugated antibodies are antibodies that bind at least one effector molecule, and are the prototypical drug for ADCs. Total antibody, conjugated antibody and free drug are key analytes required to establish exposure-response relationships for ADCs. Therefore, quantification of conjugated antibodies is crucial in pharmacokinetic studies of ADCs, as it provides critical information on the distribution, metabolism, and elimination of ADCs in vivo.
Conjugated Antibody Quantification Strategy at Creative Proteomics
Creative Proteomics' characterization team is well versed in a variety of ADCs bioanalytical techniques and is able to employ customized assay strategies to support our clients' demands. Ligand binding assay (LBA) using an anti-payload antibody is our preferred method for conjugated antibody detection of ADCs. In addition, a 'hybrid' assay combining immunoaffinity enrichment and LC-MS analysis also is widely used in our laboratory. In this hybrid assay format, our assay experts enrich samples by immunocapture with anti-payload antibodies that are digested, and then quantify the antibodies using a bottom-up LC-MS method.
| Methods |
Advantages |
| Ligand Binding Assay |
- Highly sensitive and efficient
- Gold standard for quantification of large molecules in biological samples
- Cost-effective testing
- High throughput
- No purification step required
|
| LC-MS Assay |
- Highly specific
- Resistant to interferences
- Less stringent requirement for reagents
- Potential for multiplexed analysis for multiple analytes
- Provides ADC analyte structure information
|
Advantages of Our Conjugated Antibody Quantification Services
- A wide range of anti-payload reagents are available.
- Specialized assay teams can easily meet the challenges in the hybrid assay format of LBA and LC-MS.
- Selection of signature peptides for antibody quantitation based on sample characteristics of ADCs, especially for those ADCs with non-cleavable linkers.
Creative Proteomics' characterization experts are well versed in all bioanalytical strategies for ADCs and are committed to providing our customers with highly sensitive assays to best support the analysis of their ADCs development programs. Contact us to learn more about our service and we will be happy to serve you.
