Determination of residual dextran sulfate in protein products by SEC-HPLC
| Autor: | Loubna M. Tazi, Shiranthi Jayawickreme |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Clinical Biochemistry 01 natural sciences Biochemistry Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Limit of Detection Trichloroacetic acid Chromatography High Pressure Liquid High concentration Chromatography Sec hplc 010401 analytical chemistry Metachromasia Dextran Sulfate Proteins Reproducibility of Results Cell Biology General Medicine 0104 chemical sciences 030104 developmental biology Dextran Dextran sulfate chemistry Cell culture Chromatography Gel Linear Models Molar mass distribution |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 1011 |
| ISSN: | 1873-376X |
| Popis: | Dextran sulfate is a polyanionic derivative of dextran, produced by esterification of dextran with chlorosulphonic acid. Dextran sulfate with an average molecular weight of 8000Da can be added to the cell culture to inhibit binding of proteins to cells, increasing cellular growth and productivity. Residual dextran sulfate levels must be monitored during the purification process development to insure clearance. A size-exclusion chromatography based HPLC assay has been developed for the separation and quantitation of dextran sulfate in a highly concentrated purified protein drug substance sample. Trichloroacetic acid (TCA) was used to precipitate the protein and separate the dextran sulfate. Detection and quantitation of dextran sulfate was achieved by post column reaction with dimethylene blue to form a metachromatic complex that absorbs visible light at 530nm. The quantitation limit (LOQ) was determined to be 1.5μg/mL dextran sulfate in high concentration protein samples. |
| Databáze: | OpenAIRE |
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