Domain antibodies (dAbs) are the smallest functional binding units of antibodies, corresponding to the variable regions of either the heavy (VH) or light (VL) chains of human antibodies. Different dAbs can show different degrees of self-association, which drives the need for a cheap, quick — yet robust — technique to study self-association behaviour in drug discovery. This study outlines the use of SEC–MALS to determine self-association, and validates the work by comparison with analytical ultracentrifugation (AUC).
Shimazdu LC-20AD Prominence was used in series with miniDAWN TREOS and Optilab REX for this study. The columns used were TSK-GEL3000SWXL and Superdex S200.
While AUC is the gold standard for the determination of self-association in solution, scientists are looking for alternatives as a result of AUC's high costs, low throughput, and expertise requirements. Here, SEC–MALS has shown excellent agreement with AUC results and can be used as a cheaper, easier, and quicker alternative.
This note was graciously submited by M. Burman and O. Schon, GlaxoSmithKline, 315 Cambridge Science Park, Cambridge CB4 0WG, United Kingdom.
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