Purification of therapeutic adenoviruses using laterally-fed membrane chromatography
| Autor: | Raja Ghosh, Karina Kawka, Umatheny Umatheva, Pedram Madadkar, Brian D. Lichty, Maria Fe C. Medina, Shabnam Shoaebargh, David R. Latulippe |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Chromatography
Resolution (mass spectrometry) Chemistry Filtration and Separation 02 engineering and technology 010402 general chemistry 021001 nanoscience & nanotechnology 01 natural sciences Biochemistry 0104 chemical sciences Membrane Distribution pattern Protein purification Stepwise elution Gradient elution General Materials Science Radial flow Physical and Theoretical Chemistry 0210 nano-technology Total protein |
| Zdroj: | Journal of Membrane Science. 579:351-358 |
| ISSN: | 0376-7388 |
| Popis: | Viruses are an emerging class of biotherapeutics with the potential to treat cancer, genetic, cardiovascular and other diseases. Their commercial success depends on cost-effective purification processes. Membrane chromatography processes offer great potential for virus purification, however conventional devices are known to suffer from issues related to resolution and sample dilution. Laterally-fed membrane chromatography (LFMC) overcomes these issues by minimizing the dead-volume and establishing a uniform flow distribution pattern within the device. While the efficiency of LFMC has been previously demonstrated for protein purification studies, here we present a direct comparison of the performance of a LFMC device and a conventional radial flow device (both containing 1 mL of the same strong anion-exchange membrane) for the purification of adenoviruses. Using a gradient elution strategy, a 74% virus recovery with 4% of residual amount of total protein and 1% residual amount of total DNA was obtained using the LFMC device; for the radial flow device, only 20% virus recovery was achieved for a similar level of purity. Using a stepwise elution strategy, close to 100% virus recovery with 10% residual protein and DNA was achieved with both devices. However, the product was at least 50% more dilute with the radial flow device. Overall, this study demonstrates the considerable benefits of using LFMC over conventional membrane chromatography devices for purifying virus-based biotherapeutics. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect