การศึกษาจลนศาสตร์ในการสังเคราะห์ Recombinant NS1 protein ของไวรัสแดงกี่โดยเซลล์แมลง

Name: อริสรา เอี่ยมสืบทับ

Organization : มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

keyword: Baculovirus

; Sf-9 cells

; Recombinant

Abstract: Recombinant dengue viral (NS1) protein was produced from Baculovirus Expression Vector System(BEVS). Insect cells were used as hosts for the production of recombinant NS1 protein wheninfected with recombinant NS1 baculovirus that had been engineered by insertion of NS1 gene intothe baculovirus genome. This study focused on the kinetic of recombinant dengue viral (NS1)protein synthesis by insect cell culture. The growth and nutrient consumption and by-productsaccumulation of uninfected and infected Sf-9 cells were examined. The results from the study of Sf-9 culture showed that the maximum cell density and specific growth rate of Sf-9 cells were 1.1x107cells/ml and 0.031 h-1, respectively. Glucose depleted during the culture period and the limitation ofglucose correlated with decreasing of viable cell density. Amino acids were not limited throughoutcell growth and glutamine was consumed at the highest rate. Alanine, lactate and ammonia werefound to be accumulated to approximately 12, 4.3 and 27 mmol/L, respectively.Sf-9 cells were infected with various amounts of recombinant NS1 baculovirus (MOI 0.5, 1, 5 and10). The nutrient consumptions and by-products accumulations of infected cell culture wereexamined. The results showed that unlike non infected Sf-9 cells, glucose and glutamine in infectedSf-9 cell culture were not limited because the growth rate and nutrient consumption rate of infectedSf-9 cells were lower than uninfected Sf-9 cells. Glutamine was the highest consumed amino acid.The alanine, lactate and ammonia accumulation rates were lower than uninfected culture anddecrease with the increasing multiplicity of infection (MOI) of recombinant NS1 baculovirus.Maximum yield (99 mg/L) of recombinant dengue viral NS1 protein was obtained from the cellsinfected with MOI 1 at 7 days post infection.The information obtained from this study can be used for recombinant NS1 protein production suchas the optimal amount of recombinant baculovirus used to infect into Sf-9 cells growing at earlyexponential phase was at MOI 1 and this information can used for design the low cost formula ofmedium for culturing Sf-9 cells such as lower amounts of amino acids in medium can be used.

มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: ดร.เพ็ญจันทร์ เมฆวิจิตรแสง

Role: อาจารย์ที่ปรึกษา

Created: 2546

Issued: 2005-11-08

Modified: 2006-05-09

วิทยานิพนธ์/Thesis

ISBN: 9741883121

CallNumber: BIT470

tha

DegreeName: วิทยาศาสตรมหาบัณฑิต

Level: ปริญญาโท

Descipline: เทคโนโลยีชีวภาพ

Grantor: มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

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