期刊
JOURNAL OF BIOTECHNOLOGY
卷 148, 期 4, 页码 171-181出版社
ELSEVIER
DOI: 10.1016/j.jbiotec.2010.05.005
关键词
Surface plasmon resonance; Ion-exchange chromatography; Scaled-down model for process development; Predictive modeling; Baculovirus; Biopharmaceutical
资金
- European Commission [LSHB-2006-037541, LSHB-2006-018933]
- Portuguese Fundacao para a Ciencia e Tecnologia [PTDC/EQU-EQU/71645/2006, SFRH/BD/31257/2006]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/31257/2006, PTDC/EQU-EQU/71645/2006] Funding Source: FCT
The binding and elution of the key components of a bioreaction bulk for the production of recombinant baculoviruses-a promising viral vector for gene therapy and vaccination on a model ion-exchange surface have been successfully measured and interpreted by surface plasmon resonance (SPR) spectroscopy. The micro-scaled, ion-exchange surface was produced by immobilizing a typical ion-exchange ligand, diethylaminoethyl, onto commercially available planar gold sensor chip surfaces, which were pre-derivatized with a self-assembled monolayer of 11-mercaptoundecanoic acid. Each isolated analyte was injected into the SPR cell at defined operating conditions of salt and solute concentrations to determine the adsorption equilibrium plateau, and then eluted at the same salt concentration, upon which a well-defined, residual desorption equilibrium plateau was observed. From the analysis of the binding and elution curves and equilibrium plateaus for seven key biomolecules, it is possible to determine the adsorption isotherms over a broad range of equilibrium conditions for the three main cuts of the baculovirus bioreaction bulk: the product (the infective baculovirus), the main product-related impurities, and the main process-related impurities. A model that quantitatively explains the SPR-derived adsorption/desorption data was successfully developed for this complex biological system. (C) 2010 Elsevier B.V. All rights reserved.
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