SEC

Size exclusion chromatographySize exclusion chromatography (SEC) is a chromatographic separation technique were molecules and particles are separated based on size. As stationary phase sepharose beads are used, which are spheres with a diameter between 60 to 200 µm. These beads contain pores through which molecules can penetrate in function of their size. Small molecules will elute later because they can penetrate through several pores. This in contrast to the molecules with a bigger size that can not penetrate easily through the pores. In the LECR, we make in-house SEC columns by using 10 mL syringes loaded with Sepharose CL-2B. These are used for EV isolation, separating EVs from the bulk of proteins present in the biological matrix.

 

 

Reference: Vergauwen, G., et al., Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research. Scientific Reports, 7(1), 12, 2017