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Determination of Drug-to-Antibody Ratio (DAR) by UV-Vis Spectroscopy (CAT#: STEM-MB-0893-WXH)

Introduction

Drug-to-antibody ratio (DAR) is the average number of drugs conjugated to the antibodies, which is an important attribute of ADCs. The DAR value affects the efficacy of the drug, as low drug loading reduces the potency, while high drug loading can negatively affect pharmacokinetics (PK)1 and toxicity.




Principle

UV-Vis spectroscopy is an analytical technique that measures the amount of discrete wavelengths of UV or visible light that are absorbed by or transmitted through a sample in comparison to a reference or blank sample. This property is influenced by the sample composition, potentially providing information on what is in the sample and at what concentration. The only requirement is that the sample absorb in the UV-Vis region, i.e. be a chromophore. Absorption spectroscopy is complementary to fluorescence spectroscopy. Parameters of interest, besides the wavelength of measurement, are absorbance (A) or transmittance (%T) or reflectance (%R), and its change with time.

Applications

UV/Vis spectroscopy is routinely used in analytical chemistry for the quantitative determination of diverse analytes or sample, such as transition metal ions, highly conjugated organic compounds, and biological macromolecules. Spectroscopic analysis is commonly carried out in solutions but solids and gases may also be studied.

Procedure

1. Calibrate the Spectrometer
2. Perform an Absorbance Spectrum
3. Kinetics Experiments with UV-Vis Spectroscopy

Materials

UV/VIS Spectrophotometer