Analysis of the influence of five variables on an established immunoaffinity chromatography procedure to purify a picchia pastoris yeast derived-HBsAg for pharmaceutical use
Immunoaffinity chromatography (IAC) has a wide application in protein purification. The aim of the study was to investigate the influence of five operational factors on an IAC procedure to purify a yeast derived-Hepatitis B surface antigen (rHBsAg) for pharmaceutical use. The immunosorbent adsorptio...
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| Autores principales: | , , , , , , , , , , , , , |
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| Formato: | Articulo |
| Lenguaje: | Inglés |
| Publicado: |
2008
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| Materias: | |
| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/7705 http://www.latamjpharm.org/trabajos/27/6/LAJOP_27_6_1_12_LD68X28956.pdf |
| Aporte de: |
| Sumario: | Immunoaffinity chromatography (IAC) has a wide application in protein purification. The aim of the study was to investigate the influence of five operational factors on an IAC procedure to purify a yeast derived-Hepatitis B surface antigen (rHBsAg) for pharmaceutical use. The immunosorbent adsorption
capacity of the rHBsAg is affected at 4 °C. The applied antigen concentration, 100-1000 μg mL<sup>-1</sup>,
does not have influence on this IAC efficiency. The residence time, applied antigen amount and column geometry have a significant influence on the adsorption and elution capacity, and recovery of the rHBsAg. There is a marked retention of the rHBsAg into the matrix, which is reduced at residence times higher than 2 h. A high height/diameter ratio of the column, 3.75, reduces the antigen adsorption to the matrix but increases the recovery and the productivity for this high molecular weight (multiple chemical forms) antigen and a high affinity constant ligand. The solution of the rHBsAg retention into the matrix could increase by 30% the active pharmaceutical ingredient purification and the Hepatitis B vaccine production. |
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