RESUMEN
Plasmid DNA for biopharmaceutical applications is produced easily in Escherichia coli bacteria. The cell lysis is the most crucial step for purification of plasmid DNA. In this paper, we describe a continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK-HGF using hollow fiber ultrafiltration column as a lysis chamber and compare the plasmid DNA yield and homogeneity with the T-connector and manual processes, respectively. The results show that the plasmid pUDK-HGF yield of the combination process is 13% higher than manual lysis, twice higher than using T-connector. When the proportion of lysed cells and neutralization solution is 3:1, the plasmid pUDK-HGF yield can improve by 70%. This process could be easily scaled up to meet the industrial scale for cell lysis.
Asunto(s)
Álcalis/química , Escherichia coli/citología , Escherichia coli/genética , Plásmidos/aislamiento & purificación , Biotecnología , FermentaciónRESUMEN
OBJECTIVE@#To evaluate the clinical therapeutic effect of Rabdosia rubescens aqueous extract on chronic pharyngitis and its safety.@*METHODS@#One hundred and two patients suffered from chronic pharyngtis were randomly divided into an interventional group (n=51) and a placebo group (n=51). These volunteers were given 220 mL test materials once a day double-blindly for 30 days. All patients took routine physical examinations, and their throat symptom and pharynx signs were observed before and after the experiment.@*RESULTS@#After the patients had consumed Rabdosia rubescens aqueous extract and the placebo for 30 days, the improvement rates of main clinical symptom and signs in the interventional group were 67.31% and 61.54%, while those in placebo group were 19.23% and 11.54%, with significant difference (P0.05). But after 30 days, the integral of main clinical symptom and signs in the interventional group significantly decreased (P<0.05), compared with itself before the experiment and the placebo group.@*CONCLUSION@#Rabdosia rubescens aqueous extract is safe and effective for patients with chronic pharyngitis.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedad Crónica , Método Doble Ciego , Medicamentos Herbarios Chinos , Usos Terapéuticos , Isodon , Química , Faringitis , Quimioterapia , FitoterapiaRESUMEN
pUDK-HGF, the recombinant plasmid DNA encoding human hepatocyte growth factor (HGF), can treat ischaemic disease. A great quantity of pharmaceutical pUDK-HGF is needed. A pilot-scale production process of pUDK-HGF was established based on a new chromatographic media (plasmidselect), including fermentation, cell harvesting, alkaline lysis, ultrafiltration, RNA removing and buffer exchanging on Sephacryl S-1000, capturing supercoiled plasmid DNA with plasmidselect, and removing the salt with Sepharose 6BFF. The process does not use RNase enzyme and toxic solvents.