Physicochemical Properties Analysis of Antibody Drug Conjugates
Online Inquiry
Antibody drug conjugates (ADCs) are growing and playing an important role in therapeutic applications. Understanding the physicochemical properties of ADCs is critical to ensuring their stability, efficacy, and safety, and provides valuable insights into the discovery and development of this class of drugs. Creative Proteomics offers a wide range of characterization services utilizing our advanced platform technologies to help our clients fully understand the physicochemical properties of their antibody drug conjugates and meet all the regulatory expectations and specifications for the successful release of their ADCs drugs.
Overview of Physicochemical Characterization of Antibody Drug Conjugates (ADCs)
Monoclonal antibodies typically have complex heterogeneity (e.g., glycosylation and other post-translational modifications, etc.). The superposition of the heterogeneity of the monoclonal antibody itself and the heterogeneity caused by the coupling can lead to a significant increase in the complexity of the ADCs. For ADCs with a high degree of heterogeneity (even for products utilizing targeted coupling techniques), comprehensive analysis using reliable analytical methods with sufficient resolution is required to elucidate the diversity of product-related substances. Appropriate characterization methods should be selected to fully characterize the physicochemical properties of ADCs based on the chemical nature of the small-molecule drug and linker, the mode of coupling (amino-coupling, sulfhydryl-coupling, fixed-point coupling, etc.), and the complexity of the product.
Our Physicochemical Property Analysis Services for ADCs
Creative Proteomics provides clients with comprehensive solutions for the physicochemical analysis of ADCs. Our broad portfolio of services includes assessment of the effect of coupling process on antibody primary structure, identification of drug primary coupling sites, drug-antibody coupling ratio (DAR) and drug loading distribution, molecular size variants, charge variants, and advanced structural analysis. Our characterization experts use appropriate and advanced analytical techniques according to the characteristics of different ADCs to conduct comprehensive and detailed analyses from the perspective of physical and chemical properties to reflect the quality attributes of the final ADCs products.
We use peptide mapping, mass spectrometry and other methods to assess the effect of the coupling process on the primary structure (e.g., amino acid sequence integrity, disulfide bonding), glycosylation modifications and post-translational modifications of the monoclonal antibody. If the coupling process employed has been shown not to affect glycosylation modifications, the coupled ADC can be characterized without repeating glycosylation modification-related characterization. In addition, we used mass spectrometry to identify and analyze the major coupling sites of small molecule drugs and monoclonal antibodies in ADCs.
DAR is one of the important quality attributes of ADCs that directly affects their safety and efficacy. It represents the average number of small molecule drugs coupled to each antibody molecule. Our experienced scientists select the appropriate analytical means based on the linker, the chemical nature of the small molecule drug, and the mode of coupling. Commonly used methods include ultraviolet-visible spectrophotometry (UV), hydrophobic chromatography (HIC-HPLC), reversed-phase chromatography (RP-HPLC), and mass spectrometry (MS).
- Drug loading distribution
ADCs, especially ADCs that employ non-site-specific coupling, usually contain a mixture of ADC molecules with different amounts of small molecule drugs attached to them. The drug load distribution represents the proportion of ADC molecules coupled with different amounts of small molecule drugs to the total drug molecules. In the Creative Proteomics laboratory, we use appropriate analytical methods such as hydrophobic high-performance liquid chromatography (HIC-HPLC), reversed-phase high-performance liquid chromatography (RP-HPLC), capillary electrophoresis (CE), or mass spectrometry (MS) to characterize the distribution of different drug-loading components.
We use a range of suitable methods such as Size exclusion chromatography (SEC-HPLC), non-reduced and reduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), sodium dodecyl sulfate-capillary gel electrophoresis (CE-SDS), and analytical ultracentrifugation (AUC), among many other state-of-the-art techniques, to molecular size variants (i.e., polymers and fragments) of ADCs for accurate characterization. We are particularly concerned about aggregates because many small molecule drugs coupled to antibodies are highly hydrophobic and may increase the formation of aggregates during production and storage.
Common methods offered by Creative Proteomics for charge variant determination include Capillary Zone Electrophoresis (CZE), Ion Exchange Chromatography (IEX-HPLC), Capillary Isoelectric Focusing Electrophoresis (CIEF) or Imaging Capillary Isoelectric Focusing Electrophoresis (iCIEF). Our experienced scientists select the most appropriate analytical method for our clients to characterize the charge heterogeneity of ADCs based on the properties of the linker-small molecule drug (especially charge) and the choice of coupling sites (e.g., lysine, interchain sulfhydryl groups, carbohydrates, etc.).
At Creative Proteomics, we confirm the high-level structure of ADCs through biological activity assays in addition to directly assessing the covalent structure of ADC molecules. We use ELISA, surface plasmon resonance and other methods to analyze ADCs in a comprehensive and detailed manner.
Advantages of Our Physicochemical Property Analysis Services for ADCs
- Flexible and customized characterization solutions based on the unique characteristics of different antibody drug conjugates.
- Specialized techniques and equipment designed specifically for the analysis of antibody drug conjugates.
- Adherence to quality standards and regulatory guidelines to provide accurate and reliable analytical results.
Creative Proteomics understands the importance of these characterization studies and offers a comprehensive suite of services to support the development and analysis of antibody-coupled drugs. Contact us to learn more about our service and we will be happy to serve you.
