Differential elution of Clq, Clr and Cls from human Cl bound to immune aggregates. Use in the rapid purification of Cl subcomponents
| Autor: | Maurice G. Colomb, Gérard J. Arlaud, Anne-Marie Duplaa, Robert B. Sim |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Gel electrophoresis
Chromatography Elution Ovalbumin Immunology Osmolar Concentration chemistry.chemical_element Calcium Hydrogen-Ion Concentration Immunodiffusion Electrophoresis chemistry Ionic strength Complement C1 Yield (chemistry) Immunoglobulin G Humans Electrophoresis Polyacrylamide Gel Complement Pathway Classical Molecular Biology Polyacrylamide gel electrophoresis Protein Binding |
| Zdroj: | Molecular immunology. 16(7) |
| ISSN: | 0161-5890 |
| Popis: | IgG-ovalbumin aggregates were used to bind and activate Cl from human serum. The resulting Cl bound to the insoluble support provided a convenient tool for studying the release of Cl sub-components under various conditions of pH and ionic strength. In the presence of calcium, Clr and Cls were removed in parallel under all the conditions employed, with a minimum release at pH 7.0 and at low ionic strength. The elution behaviour of Clq was distinct from that of the Clr-Cls pair. The experimental conditions used demonstrated the presence of two separate entities in Cl: firstly, Clq and secondly, Clr plus Cls. Cl bound to IgG-ovalbumin was employed for a rapid purification of Clq, Clr and Cls sub-components. Clr plus Cls were first selectively released with EDTA † . Clr was separated from Cls by DEAE-cellulose chromatography, and Cls was further purified on anti-Clr IgG-Sepharose 6B in order to remove contaminant Clr. Clq still bound to IgG-ovalbumin aggregates was removed at pH 10.0 in the presence of 0.7 M NaCl and further purified by CM-cellulose chromatography. Clq, Clr and Cls were obtained in good yield and in pure form, as judged by immunodiffusion analysis and SDS-polyacrylamide gel electrophoresis. |
| Databáze: | OpenAIRE |
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