Antibody Drug Conjugate (ADC) Analysis

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There has been a significant resurgence in the development of antibody-drug conjugates (ADC) as target-directed therapeutic agents for cancer treatment. Among the factors critical to effective ADC design is the Drug Antibody Ratio (DAR). The DAR describes the degree of drug addition which directly impacts both potency and potential toxicity of the therapeutic, and can have significant effects on properties such as stability and aggregation. Determination of DAR is, therefore, of critical importance in the development of novel ADC
therapeutics.

DAR is typically assessed by mass spectrometry (MALDI-TOF or ESI-MS) or UV spectroscopy. Calculations based on UV absorption are often complicated by similarities in extinction coefficients of the antibody and small molecule. Mass spectrometry, though a powerful tool for Mw determination, depends on uniform ionization and recovery between compounds—which is not always the case for ADCs.

We present here a method for DAR determination based on SEC-MALS in conjunction with UV absorption and differential refractive index detection.

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