Preparation and testing of stationary phases and modified capillaries for affinity chromatography and affinity capillary electrophoresis of pepsin
Language English Country Netherlands Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
16114256
DOI
10.1016/j.chroma.2004.10.022
PII: S0021-9673(04)01820-5
Knihovny.cz E-resources
- MeSH
- Chromatography, Affinity instrumentation methods MeSH
- Diiodotyrosine chemistry MeSH
- Electrophoresis, Capillary instrumentation methods MeSH
- Humans MeSH
- Silicon Dioxide chemistry MeSH
- Pepsin A isolation & purification MeSH
- Swine MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Diiodotyrosine MeSH
- Silicon Dioxide MeSH
- Pepsin A MeSH
Three stationary phases have been prepared for affinity liquid chromatography isolation and separation of porcine and human pepsin. The phases contain 3,5-diiodo-L-tyrosine (DIT) bound to the supports HEMA BIO VS, HEMA BIO E and EPOXY TOYOPEARL. These phases have been tested on a model sample of porcine pepsin A and applied to human pepsin. Fractions have been collected and the chymase activity determined in selected analyses. For affinity CE, capillaries have been prepared by modifying the wall with 3-aminopropyltriethoxysilane, followed either by direct binding of DIT, or by binding L-tyrosine that was subsequently iodated. The dissociation constant K(d) has been determined for the pepsin-DIT complex from the changes in the electrophoretic mobilities.
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