In this study, the CaP/pDNA nanoparticles were prepared using Triton X-100/Butanol/Cyclohexane/Water reverse microemulsion system. Optimization of preparation conditions was based on evaluation of particle size by Box...In this study, the CaP/pDNA nanoparticles were prepared using Triton X-100/Butanol/Cyclohexane/Water reverse microemulsion system. Optimization of preparation conditions was based on evaluation of particle size by Box–Behnken design method. The particle sizes of the optimized CaP/pDNA nanoparticles were found to be 60.23 ± 4.72 nm, polydispersity index was 0.252 and pDNA encapsulate efficiency was more than 90%. The optimized CaP/pDNA nanoparticles have pH sensitivity and biocompatibility. Further, optimized CaP/pDNA nanoparticles showed higher transfection efficiency.展开更多
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
Inherited genetic disorders of the liver pose a significant public health burden.Liver transplantation is often limited by the availability of donor livers and the exorbitant costs of immunosuppressive therapy.To over...Inherited genetic disorders of the liver pose a significant public health burden.Liver transplantation is often limited by the availability of donor livers and the exorbitant costs of immunosuppressive therapy.To overcome these limitations,nucleic acid therapy provides a hopeful alternative that enables gene repair,gene supplementation,and gene silencing with suitable vectors.Though viral vectors are the most efficient and preferred for gene therapy,pre-existing immunity debilitating immune responses limit their use.As a potential alternative,lipid nanoparticle-mediated vectors are being explored to deliver multiple nucleic acid forms,including pDNA,mRNA,siRNA,and proteins.Herein,we discuss the broader applications of lipid nanoparticles,from protein replacement therapy to restoring the disease mechanism through nucleic acid delivery and gene editing,as well as multiple preclinical and clinical studies as a potential alternative to liver transplantation.展开更多
Developing new drugs to treat Parkinson's disease efficiently is challenging. Here we report that chitosan nanoparticles(APPDNs) could serve as novel candidates for the design of anti-PD drugs. In this study, we i...Developing new drugs to treat Parkinson's disease efficiently is challenging. Here we report that chitosan nanoparticles(APPDNs) could serve as novel candidates for the design of anti-PD drugs. In this study, we investigated the effects of chitosan poly ethyleneglycol-poly lactic acid(PEG-PLA) nanoparticles conjugated with nerve growth factor(NGF), acteoside(ACT) and plasmid DNA(p DNA) for PD therapy using in vitro and in vivo models. Using PD cell models, we demonstrated that APPDN had good neuroprotective effects. More significantly, experiments using mouse PD models demonstrated that APPDNs could ameliorate the behavioral disorders of sick mice. Immunohistochemical and western blot(WB) analyses demonstrated that APPDNs could significantly reverse dopaminergic(DA) neuron loss in the substantia nigra and striatum of sick mice. This study opens up a novel avenue to develop anti-PD drugs.展开更多
基金the Natural Science Foundation Committee of China for the financial support (No.81173004 and No.81202483)
文摘In this study, the CaP/pDNA nanoparticles were prepared using Triton X-100/Butanol/Cyclohexane/Water reverse microemulsion system. Optimization of preparation conditions was based on evaluation of particle size by Box–Behnken design method. The particle sizes of the optimized CaP/pDNA nanoparticles were found to be 60.23 ± 4.72 nm, polydispersity index was 0.252 and pDNA encapsulate efficiency was more than 90%. The optimized CaP/pDNA nanoparticles have pH sensitivity and biocompatibility. Further, optimized CaP/pDNA nanoparticles showed higher transfection efficiency.
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.
基金the Department of Biotechnology,India,for the financial support through grants,BT/PR25841/GET/119/162/2017,BT/PR40446/COV/140/5/2021。
文摘Inherited genetic disorders of the liver pose a significant public health burden.Liver transplantation is often limited by the availability of donor livers and the exorbitant costs of immunosuppressive therapy.To overcome these limitations,nucleic acid therapy provides a hopeful alternative that enables gene repair,gene supplementation,and gene silencing with suitable vectors.Though viral vectors are the most efficient and preferred for gene therapy,pre-existing immunity debilitating immune responses limit their use.As a potential alternative,lipid nanoparticle-mediated vectors are being explored to deliver multiple nucleic acid forms,including pDNA,mRNA,siRNA,and proteins.Herein,we discuss the broader applications of lipid nanoparticles,from protein replacement therapy to restoring the disease mechanism through nucleic acid delivery and gene editing,as well as multiple preclinical and clinical studies as a potential alternative to liver transplantation.
基金supported financially by the National Natural Science Foundation of China(Nos.31370967 and 31170919)the Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme(2014)+2 种基金the Science and Technology Planning Project of Guangdong Province(No.2015A020212033)the Science and Technology Project of Guangzhou(No.201805010002)the Innovation Project of Graduate School of South China Normal University(No.2018LKXM019)。
文摘Developing new drugs to treat Parkinson's disease efficiently is challenging. Here we report that chitosan nanoparticles(APPDNs) could serve as novel candidates for the design of anti-PD drugs. In this study, we investigated the effects of chitosan poly ethyleneglycol-poly lactic acid(PEG-PLA) nanoparticles conjugated with nerve growth factor(NGF), acteoside(ACT) and plasmid DNA(p DNA) for PD therapy using in vitro and in vivo models. Using PD cell models, we demonstrated that APPDN had good neuroprotective effects. More significantly, experiments using mouse PD models demonstrated that APPDNs could ameliorate the behavioral disorders of sick mice. Immunohistochemical and western blot(WB) analyses demonstrated that APPDNs could significantly reverse dopaminergic(DA) neuron loss in the substantia nigra and striatum of sick mice. This study opens up a novel avenue to develop anti-PD drugs.
