Displacement chromatography of proteins using a self-sharpening pH front formed by adsorbed buffering species as the displacer
Narahari, CR; Strong, JC; Frey, DD
| HERO ID | 5163292 |
|---|---|
| In Press | No |
| Year | 1998 |
| Title | Displacement chromatography of proteins using a self-sharpening pH front formed by adsorbed buffering species as the displacer |
| Authors | Narahari, CR; Strong, JC; Frey, DD |
| Journal | Journal of Chromatography A |
| Volume | 825 |
| Issue | 2 |
| Page Numbers | 115-126 |
| Abstract | 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 beta-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 x 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. |
| Doi | 10.1016/S0021-9673(98)00718-3 |
| Pmid | 9842719 |
| Wosid | WOS:000077119900002 |
| Is Certified Translation | No |
| Dupe Override | No |
| Is Public | Yes |
| Language Text | English |
| Keyword | displacement chromatography; preparative chromatography; chromatofocusing; mobile phase composition; proteins; lactoglobulins |