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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Zhou, Ling Danielson, Neil D. |
| Description | Author Affiliation: Zhou L ( Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, United States. Electronic address: zhoul3@miamioh.edu.) |
| Abstract | The room temperature ionic liquid isopropylammonium formate (IPAF) is studied as a reversed phase HPLC mobile phase modifier for separation of native proteins using a polymeric column and the protein stability is compared to that using acetonitrile (MeCN) as the standard organic mobile phase modifier. A variety of important proteins with different numbers of subunits are investigated, including non-subunit proteins: albumin, and amyloglucosidase (AMY); a two subunit protein: thyroglobulin (THY); and four subunit proteins: glutamate dehydrogenase (GDH) and lactate dehydrogenase (LDH). A significant enhancement in protein stability is observed in the chromatograms upon using IPAF as a mobile phase modifier. The first sharper peak at about 2min represented protein in primarily the native form and a second broader peak more retained at about 5-6min represented substantially denatured or possibly aggregated protein. The investigated proteins (except LDH) could maintain the native form within up to 50% IPAF, while a mobile phase, with as low as 10% MeCN, induced protein denaturation. The assay for pyruvate using LDH has further shown that enzymatic activity can be maintained up to 30% IPAF in water in contrast to no activity using 30% MeCN. |
| File Format | HTM / HTML |
| ISSN | 15700232 |
| e-ISSN | 1873376X |
| DOI | 10.1016/j.jchromb.2013.08.005 |
| Journal | Journal of Chromatography B |
| Volume Number | 940 |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2013-12-01 |
| Publisher Place | Netherlands |
| Access Restriction | Open |
| Subject Keyword | Discipline Analytical Chemistry Chromatography, Reverse-phase Formic Acid Esters Chemistry Ionic Liquids Proteins Acetonitriles Chromatography, High Pressure Liquid Models, Chemical Propylamines Protein Stability Protein Subunits Isolation & Purification Quaternary Ammonium Compounds Research Support, N.i.h., Extramural |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry |
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