Can you separate small molecules & peptides or large molecules the same way? | Trust Your Science 16

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December 7, 2018

Many laboratories today are transitioning from analyzing small molecules to large molecules. A chromatography analyst asks #TrustYourScience: I’ve heard you have to treat peptides differently than small molecules in a chromatography workflow? Is that true? Small molecules are usually less than 1 kDa molecular weight. Peptides are usually around 4 kDa. Monoclonal antibodies, or proteins, can be around 150 kDa, so they’re going to behave differently in chromatographic separations. Jon and Kim pick a column particle side – 3.5 microns – and run a variety of compounds on it with different molecular weights. They use the Van Deemter curves to determine optimal flow rates, and to see if they have to treat large molecules differently than small molecules. They also check if column particle size makes a difference.

Analytical TechniquesChromatographyMass Spectrometry

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