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Preliminary study on the optimization of recombinant human transferrin expression in insect cells baculovirus system

机译：昆虫细胞杆状病毒系统中重组人转铁蛋白表达优化的初步研究

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摘要

Insect cells Baculovirus system is a promising artificial system for producing many biopharmaceuticals products. In this study, the expression profiles of recombinant human transferrin (rhTf), a model protein expressed in Sf9 insect cells monolayer and baculovirus culture were investigated. Time course expression profile of rhTf at various multiplicity of infection (MOI), seeding density (SD), time of infection (TOI), and harvest time (HT) were conducted as well as growth control (GC) for non infected Sf9 cells. Growth kinetics of non infected and infected Sf9 cells were also performed to monitor the progress of infection. SDS-PAGE, Western Blot, and ELISA analyses were utilized in this study to confirm the progress of the rhTf production. The data shows that maximum production of rhTf was between 24 to 72 hours post infection, at the seeding density of 1.6 x 106 viable cells/ml, infection at day 2 post culture, and MOI of 5. The maximum rhTf yield was 11.2 g/ml. The results suggested that rhTf yield could be increased further provided that the nutrient and cells density are not limiting.
机译：昆虫细胞杆状病毒系统是一种有前途的人工系统，可用于生产许多生物制药产品。在这项研究中，研究了重组人转铁蛋白（rhTf），在Sf9昆虫细胞单层和杆状病毒培养物中表达的模型蛋白的表达谱。对未感染的Sf9细胞进行了rhTf在各种感染复数（MOI），接种密度（SD），感染时间（TOI）和收获时间（HT）的时程表达谱以及生长控制（GC）。还进行了未感染和感染的Sf9细胞的生长动力学，以监测感染的进程。 SDS-PAGE，Western Blot和ELISA分析用于这项研究，以确认rhTf产生的进展。数据显示，rhTf的最大产量在感染后24至72小时之间，接种密度为1.6 x 106个活细胞/ ml，培养后第2天感染，MOI为5。最大rhTf产量为11.2％。克/毫升结果表明，只要营养和细胞密度不受限制，rhTf产量就可以进一步提高。

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