Displacement chromatography of proteins using a self-sharpening pH front formed by adsorbed buffering species as the displacer
| Autor: | Douglas D. Frey, Chittoor R. Narahari, John Strong |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Chromatography
Chemistry Chromatofocusing Organic Chemistry Ion chromatography Analytical chemistry Proteins Lactoglobulins General Medicine Hydrogen-Ion Concentration Chromatography Ion Exchange Biochemistry Displacement chromatography Analytical Chemistry MOPS chemistry.chemical_compound Acetic acid Adsorption Desorption Spectrophotometry Ultraviolet Ethanesulfonic acid |
| Zdroj: | Journal of Chromatography A. 825:115-126 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/s0021-9673(98)00718-3 |
| Popis: | The preparative-scale separation of two proteins into adjoined, pure bands was accomplished using a novel, hybrid chromatography method which employs chromatofocusing using a self-sharpening pH front and displacement development. The method eliminates the use of a traditional displacer for accomplishing displacement chromatography, and was used to separate the A and B forms of β-lactoglobulin using a strong-base anion-exchange column packing and a buffer system composed of acetic acid and either 3-(N-morpholino)propane-sulfonic acid (MOPS) or 2-(N-morpholino)ethanesulfonic acid (MES). Sample loads up to 150 mg of protein were applied to a 75×7.5 mm column to produce a displacement train composed of highly pure protein bands with greater than 90% recovery of protein. A discussion is given of the chromatographic behavior of proteins under concentration overloaded conditions for the case where a self-sharpening pH front formed using adsorbed buffering species is used to desorb proteins from an anion-exchange column packing. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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