สืบค้นงานวิจัย
Optimization of Recombinant Hemagglutinin Protein Production from Baculovirus Expression Vector System
Irisa Trianti - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Optimization of Recombinant Hemagglutinin Protein Production from Baculovirus Expression Vector System
ผู้แต่ง / หัวหน้าโครงการ (EN): Irisa Trianti
บทคัดย่อ (EN): Hemagglutinin (HA), the major influenza viral surface glycoprotein, could potentially be used as influenza vaccine. Baculovirus Expression Vector System (BEVS) which has been known to produce high yield of recombinant protein with biological activity, antigenicity and immunogenecity, the same as that of the natural protein, was therefore employed to produce recombinant HA protein for this purpose. The recombinant HA protein can be achieved after infection of genetically engineered baculovirus harbouring HA gene into its insect cell host. High level of recombinant HA protein production can be accomplished by optimization of several factors such as ratio of cells and viruses (MOl), time of harvest (TOH), etc. It was found that high recombinant HA protein production could be attained by infecting Sf-9 cells at the density of lx106 cells/ml with recombinant HA baculovirus at MOl of 1. In addition, recombinant HA protein production may be further optimized by reducing cell stress caused by baculovirus infection using anti-stress agent. Therefore, supplementation of anti-stress agents, catalase and selenium in the form of either sodium selenate (Na2SeO4) or sodium selenite (Na2SeO3), into infected cell culture was proposed to enhance the recombinant HA protein production. Result showed that recombinant HA protein production was increased 1.3 times in comparison with that of the control at 3 days post infection when 0.625 ng/ml of catalase was supplemented into growth medium. Moreover, when either Na2SeO3 or Na2SeO4 at the concentration of 0.4 and 0.2 ng/ml. respectively, was individually supplemented into growth medium, an increase in recombinant HA protein produced of approximately 2.4 and 2.3 fold higher than that of control at day 4 post infection, respectively, could be anticipated. It is worthy to note that addition of these anti-stress agents not only increased level of recombinant HA protein production but also shortened the production period from 5 days to 3 days (for catalase) and 4 days (for selenium). Effect of anti-stress agents on recombinant HA protein production was attained through response surface methodology, Results showed that Na2SeO4 was considered statistically significant (p<0.05), whereas effect of Na2SeO3 was subtle (p = 0.065). Additionally, supplementation of catalase appeared to play insignificant role on recombinant HA protein production. Further, it is interesting to note that interactions among these anti-stress agents, i.e., Na2SeO3 x Na2SeO4, were all negative excluding catalase x catalase, indicating their negative effects on recombinant HA protein production. Result further demonstrated that optimal concentrations of Na2SeO3 and Na2SeO4 leading to high recombinant HA protein production are, respectively, 0.18 and 0.37 ng/ml. Validation experiment conducted further showed that a slight increase in recombinant HA protein production (1.3 folds higher than that of control) could be anticipated on day 4 post infection providing the fact that, even on day 5 post infection, level of recombinant HA protein produced was approximately 1.2 folds of that of control which is better than that obtained when individual anti-stress agents was supplemented into the growth medium.
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): http://dcms.thailis.or.th/dcms/dccheck.php?Int_code=54&RecId=11007&obj_id=32765
เผยแพร่โดย (EN): มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี
คำสำคัญ (EN): Recombinant Hemagglutinin (rHA) Protein
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี
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Irisa Trianti. "Optimization of Recombinant Hemagglutinin Protein Production from Baculovirus Expression Vector System". ไม่ระบุหน่วยงาน. ม.ป.ป.

Irisa Trianti. "Optimization of Recombinant Hemagglutinin Protein Production from Baculovirus Expression Vector System". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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Optimization of Recombinant Hemagglutinin Protein Production from Baculovirus Expression Vector System
Irisa Trianti
มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี
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