High-performance size exclusion chromatography of sea worm chlorocruorin and other large proteins, viruses and polysaccharides on a TSK G5000 PW preparative column

Abstract The elution parameters of large enzymes, viruses, ribosomes and other “supramolecular” structures are studied using the preparative TSK G5000 PW type column. The pigmented protein, chlorocruorin, isolated from the sea worm Potamilla leptochaeta , was found to serve as an excellent high-molecular-weight marker for size exclusion liquid chromatography. This is due to its high degree of molecular stability and a molecular weight, found to be 2.9·10 6 by sedimentation velocity analysis, which is located in a zone formed between viruses and enzymes that is largely devoid of macromolecular markers. Calibration constants for this chromatography column are found for both molecular weight and molecular radii. The data found for hydrodynamic molecular radii are further extended to non-globular, swollen macromolecules, such as polysaccharides, using dextran fractions sized by alcohol precipitation.