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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | He, Chunyan Tong, Kuitang Yang, Zhiqiang |
| Description | Country affiliation: China Author Affiliation: He C ( School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, China. hechyan@hotmail.com) |
| Abstract | A downstream processing was examined for Vero cell-derived human influenza virus (H1N1) grown in serum free medium. Vero cell banks were established in serum free medium and characterized according to regulatory requirements. Serum free Vero cells were grown on Cytodex 3 microcarriers in 5L bioreactor and infected with influenza A virus (A/New Caledonia/99/55). The harvests were processed with the sequence of inactivation, clarification, anion exchange chromatography (DEAE FF), Cellufine Sulfate Chromatography (CSC) and size exclusion chromatography (Sepharose 6FF). Host cell DNA (hcDNA) was mainly removed with DEAE FF column and CSC by 40 and 223 fold, respectively. Most of Vero cell proteins were eliminated in CSC and Sepharose 6FF unit operation by about 13 fold. The overall scheme resulted in high recovery of hemagglutinin (HA) activity and the substantial removal of total protein, host protein and DNA. The total protein content and DNA content per 15 µg HA protein in final product was 89 µg and 33 pg, respectively, which complied with regulatory requirements for single strain influenza vaccines. SDS-PAGE analysis and Western blotting confirmed the purity of the final product. In conclusion, the suggested downstream process is suitable for the purification of microcarrier-based cell-derived influenza vaccine. |
| ISSN | 00219673 |
| Issue Number | 31 |
| Volume Number | 1218 |
| e-ISSN | 18733778 |
| Journal | Journal of Chromatography A |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2011-08-05 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Chromatography Methods Electrophoresis, Polyacrylamide Gel Influenza A Virus, H1n1 Subtype Isolation & Purification Virus Cultivation Animals Cercopithecus Aethiops Culture Media, Serum-free Analysis Humans Growth & Development Vero Cells Evaluation Studies Journal Article Research Support, Non-u.s. Gov't Discipline Analytical Chemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Medicine Analytical Chemistry Biochemistry |
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