The main component of the Center for Genetic Engineering and Biotechnology(CIGB)candidate vaccine against Hepatitis C virus(HCV)is the pIDKE2 plasmid.The current designed downstream process for the production of pIDKE...The main component of the Center for Genetic Engineering and Biotechnology(CIGB)candidate vaccine against Hepatitis C virus(HCV)is the pIDKE2 plasmid.The current designed downstream process for the production of pIDKE2 fulfils all regulatory requirements and renders the required quantities of pharmaceuticalgrade plasmid DNA(pDNA)with 95%purity.The advantages of this procedure include high plasmid purity and the elimination of undesirable additives,such as toxic organic extractants and animal-derived enzymes.However,yields and consequently the productivity of the process are low.Previous work demonstrated that the most critical step of the process is the reverse phase chromatography,where conventional porous particle resins are used.Therefore,to increase the process productivity,alternative technologies such as membranes and chromatographic monoliths were tested as alternative options for this critical step.Here,a comparison between the behaviors of CIM~ C4-HLD and Sartobind phenyl matrices was performed.To obtain higher productivities and purities,the dynamic binding capacities and selectivities were evaluated.The results showed that both matrices had a similar capacity for pIDKE2 plasmid,but the separation of pDNA isoforms using CIM~ technology was much better than that with Sartobind.Additionally,the optimal conditions for loading plasmid DNA on a CIMC4-HLD 800-mL monolithic column in a real production process were determined.These optimizations will allow production levels to satisfy the high plasmid consumption demanded by clinical trials.展开更多
MoS2/γ－Al2O3 ceramic composite membrane is successfully synthesized by the sol-gel method based on the inorganic salt route. The aluminum hydrate sol derived from the inorganic salt Al(NO3)3, whose transparence and ...MoS2/γ－Al2O3 ceramic composite membrane is successfully synthesized by the sol-gel method based on the inorganic salt route. The aluminum hydrate sol derived from the inorganic salt Al(NO3)3, whose transparence and viscosity are 97% and 1.2 × 10-3 Pa. s, respectively, can be formed through adjusting the ratio of the peptizing agent H+ to Al3+ to 0.3. The aluminum hydrate gel at 110℃ is amorphous in structure and is heat-treated in air at 800℃ to form γ－Al2O3. The precursor derived from the mixture solution of ammonium molybdate, thioacetamide and a reducing agent, can be transformed into crystal MoS2 under reducing condition at 800℃. MoS2/γ－Al2O3 composite membrane is an organic whole and bps no gradation from MoS2 layer to γ－Al2O3 Iayer. The separation factor a H2/H2S through the MoS2/γ－Al2O3 composite membrane increase with rising temperature and a H2/H2S at 600℃ is 4.45 higher than the theoretical separation factor produced by the ideal Knudsen diffusion.展开更多
基金Center for Genetic Engineering and Biotechnology and BIA Separations Fund
文摘The main component of the Center for Genetic Engineering and Biotechnology(CIGB)candidate vaccine against Hepatitis C virus(HCV)is the pIDKE2 plasmid.The current designed downstream process for the production of pIDKE2 fulfils all regulatory requirements and renders the required quantities of pharmaceuticalgrade plasmid DNA(pDNA)with 95%purity.The advantages of this procedure include high plasmid purity and the elimination of undesirable additives,such as toxic organic extractants and animal-derived enzymes.However,yields and consequently the productivity of the process are low.Previous work demonstrated that the most critical step of the process is the reverse phase chromatography,where conventional porous particle resins are used.Therefore,to increase the process productivity,alternative technologies such as membranes and chromatographic monoliths were tested as alternative options for this critical step.Here,a comparison between the behaviors of CIM~ C4-HLD and Sartobind phenyl matrices was performed.To obtain higher productivities and purities,the dynamic binding capacities and selectivities were evaluated.The results showed that both matrices had a similar capacity for pIDKE2 plasmid,but the separation of pDNA isoforms using CIM~ technology was much better than that with Sartobind.Additionally,the optimal conditions for loading plasmid DNA on a CIMC4-HLD 800-mL monolithic column in a real production process were determined.These optimizations will allow production levels to satisfy the high plasmid consumption demanded by clinical trials.
文摘MoS2/γ－Al2O3 ceramic composite membrane is successfully synthesized by the sol-gel method based on the inorganic salt route. The aluminum hydrate sol derived from the inorganic salt Al(NO3)3, whose transparence and viscosity are 97% and 1.2 × 10-3 Pa. s, respectively, can be formed through adjusting the ratio of the peptizing agent H+ to Al3+ to 0.3. The aluminum hydrate gel at 110℃ is amorphous in structure and is heat-treated in air at 800℃ to form γ－Al2O3. The precursor derived from the mixture solution of ammonium molybdate, thioacetamide and a reducing agent, can be transformed into crystal MoS2 under reducing condition at 800℃. MoS2/γ－Al2O3 composite membrane is an organic whole and bps no gradation from MoS2 layer to γ－Al2O3 Iayer. The separation factor a H2/H2S through the MoS2/γ－Al2O3 composite membrane increase with rising temperature and a H2/H2S at 600℃ is 4.45 higher than the theoretical separation factor produced by the ideal Knudsen diffusion.
