Efficient separation and analysis of antibody fragments and their drug conjugates

contributed by YMC | 04/24/2025

Alongside monoclonal antibodies (mAbs) and antibody- drug-conjugates (ADCs) antibody fragments and their drug conjugates find application in therapeutic treatments. These fragments bind selectively to specific antigens, mirroring the function of intact antibodies. Due to their significantly smaller size, antibody fragments require dedicated analyti- cal methods for precise characterisation and quality control.

This Application Note presents a precise method to separate a single-chain variable fragment (scFv) from its conjugated species (FDC). The separation was performed using a SEC- MS method. To achieve optimal resolution and peak shapes, a bioinert column hardware is essential. A metal-free PEEK- lined YMC-Pack Diol-120 column was selected for this analysis, ensuring high performance in the characterisation of the scFv with a molecular weight of approximately 26 kDa.

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