• Maximum efficiency in the analysis of mAbs and ADCs

    Ion chromatography (IC)

    Maximum efficiency in the analysis of mAbs and ADCs

    The YMC Application Collection for monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) serves as a helpful guide for the analytical characterisation and quality control of these complex biomolecules, covering essential chromatographic techniques. This document supports biopharmaceutical R&D by providing highly reproducible analytical methods with excellent resolution of commercially available mAbs and ADCs.

    1. Charge variant analysis

    Ion Exchange Chromatography (IEX) is widely used for the charge variant analysis of mAbs, including acidic and basic modifications. These charge variants are critical in assessing the quality and stability of mAbs, as such modifications can impact efficacy and safety. The guide includes both commonly used cation exchange methods for most commercially available mAbs with a higher isoelectric point (pI) and the less commonly used anion exchange method, which is more suitable for mAbs with a lower pI, such as the increasingly common IgG4-based mAbs currently in development. With BioPro IEX columns also a combination with mass spectrometric (MS) detection is also possible.

    2. Size variant analysis

    Size Exclusion Chromatography (SEC) is crucial for the analysis of mAb aggregates and fragments, aiding in the assessment of the molecular integrity of the antibody. This technique is particularly important for quality control of mAb products, as aggregation can affect pharmacokinetics and immunogenicity. The YMC-SEC MAB column is especially designed to separate aggregates, the intact antibody and its fragments simultaneously. Together with its compatibility with light scattering detectors YMC-SEC MAB is the first choice for mAbs.

    3. DAR and oxidation determination

    Due to the inherent hydrophobicity of antibodies, hydrophobic interaction chromatography (HIC) is valuable for analysing mAbs and ADCs, particularly for determining the drug-to-antibody ratio (DAR) in ADCs. Reliable determination of the DAR is crucial for the efficacy of the ADC. The included applications demonstrate reliable high-throughput analysis of DAR values by using a BioPro HIC HT column. In addition, the BioPro HIC Bf column is ideally suitable for the separation of oxidized and non-oxidized mAb variants.

    4. Intact and digested mAb evaluation

    Reverse-phase chromatography (RP) is an established method for analysing smaller peptides and antibody fragments. The analysis of intact mAbs requires elevated temperatures to prevent adsorption. YMC-Triart Bio C4 columns, with their wide-pore design and high thermal stability, perform reliably at temperatures up to 90 °C, making them an effective choice for these applications. Additionally, RP seamlessly integrates with MS detection, supporting high-sensitivity analyses with consistent performance.

    You can learn more about BioLC solutions by visiting YMC’s homepage or downloading the Biochromatography columns catalogue!


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