Evaluation of baculovirus expression vectors with enhanced stability in continuous cascaded insect-cell bioreactors.

摘要: Continuous protein production with baculovirus expression vectors in insect-cell bioreactors is characterized by a dramatic drop heterologous within few weeks. This mainly due to the spontaneous deletion of gene(s) from genome and/or rapid accumulation defective interfering baculoviruses (DIs). Cell culture experiments bacmid-derived showed that deletions foreign bacterial artificial chromosome (BAC) sequences readily occurred. These correlated low density homologous (repeat) regions (hrs), which are located dispersed throughout and believed act as origins viral DNA replication (oris). To test hypothesis more likely occur ori density, properties an additional hr unstable BAC were compared standard continuous cascaded bioreactor configuration. All viruses equipped green fluorescent (GFP) gene encoding classical swine fever virus E2 glycoprotein (CSFV-E2). The insertion extra vector led improved genetic stability adjacent sequences, resulting prolonged expression. maintenance appeared be dependent on orientation inserted hr. advantages utilization hrs improve for large-scale discussed.