Reading%20points%20for%20Scopes%20c%20090919 - CHEN 4820...

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10/12/2010 CHEN 4820 - Reading points from Scopes, Protein Purification, Principles and Practice. Sec 1.3 – Principles of column chromatography Basic idea of chromatography – solutes carried by a mobile phase through a column holding a sorbent (“matrix”) move at different speeds because they adsorb to different degrees. Factors affecting the degree of chromatographic separation: column length, degree of diffusion, extent of loading (“sample depth”). Typical elution methods: step or gradient. The minor importance of column length/diameter to separation (assuming uniform flow). Separation performance improves with smaller bead size. Importance of good design of column inlet and outlet to avoid nonuniform flow. Column packing basics to insure uniform flow – appropriate conc of resin slurry and packing flowrate. Typical means to detect and fractionate proteins emerging from the column – UV detector and fraction collector. Ch 4 – Separation by Precipitation
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Reading%20points%20for%20Scopes%20c%20090919 - CHEN 4820...

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