ISOLATION, CLONING AND EXPRESSION OF RECOMBINANT HUMAN FACTOR VII IN CHO CELL LINE

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HALABIAN R.; SHAGERDI ESMAEILI N.; OUDI A.; MASROURI N.; AMIRIZADEH N.; MOUSAVI HOSSEINI K.; GHARAH BAGHIAN A.; REZVAN H.; JALILI M.A.; HABIBI ROUDKENAR M.

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Journal: THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON) Year:2009 | Volume:6 | Issue:1 (22) Page(s): 1-11

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Title

ISOLATION, CLONING AND EXPRESSION OF RECOMBINANT HUMAN FACTOR VII IN CHO CELL LINE

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Keywords

HEMOPHILIAQ4

RFVIIAQ3

CHOQ3

TRANSFECTIONQ3

Abstract

Background and Objectives: Factor VII is a plasma glycoprotein that participates in the coagulation process leading to the generation of fibrin. Factor VII plays an important role in the cascade of coagulation. The aim of this study was to clone and express human recombinant factor VII in CHO cell line as an eukaryotic host cell.Materials and Methods: In this descriptive study, FVII cDNA was isolated from HepG2 cell line and cloned to pcDNA 30.1(+) vector. The constructs were transfected to CHO cell line. A cell line that permanently expressed recombinant factor VII was established. The expression of recombinant FVII was determined by RT-PCR, ELISA, SDS-PAGE and western blot analysis. Biological activity of recombinant factor VII was determined by prothrombin time assay in factor FVII-depleted plasma.Results: The results showed that FVII was successfully cloned and expressed. After 3 weeks stable cell lines were generated in the culture of the CHO cell line in the presence of geneticin. RT-PCR, ELISA, SDS-PAGE and western blot analysis results indicate the expression of FVII in the stable clones. A three- to four-fold decrease of the specific coagulant activity of rFVII was observed that indicates of rFVII being biologically active.Conclusions: Four thousands to Six thousands vials of rFVII were imported to our country having high cost for the government. Therefore, production of rFVII through recombinant DNA technology within lab scale is the first step to overcome the problems.

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APA: Copy

HALABIAN, R., SHAGERDI ESMAEILI, N., OUDI, A., MASROURI, N., AMIRIZADEH, N., MOUSAVI HOSSEINI, K., GHARAH BAGHIAN, A., REZVAN, H., JALILI, M.A., & HABIBI ROUDKENAR, M.. (2009). ISOLATION, CLONING AND EXPRESSION OF RECOMBINANT HUMAN FACTOR VII IN CHO CELL LINE. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), 6(1 (22)), 1-11. SID. https://sid.ir/paper/78338/en

Vancouver: Copy

HALABIAN R., SHAGERDI ESMAEILI N., OUDI A., MASROURI N., AMIRIZADEH N., MOUSAVI HOSSEINI K., GHARAH BAGHIAN A., REZVAN H., JALILI M.A., HABIBI ROUDKENAR M.. ISOLATION, CLONING AND EXPRESSION OF RECOMBINANT HUMAN FACTOR VII IN CHO CELL LINE. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)[Internet]. 2009;6(1 (22)):1-11. Available from: https://sid.ir/paper/78338/en

IEEE: Copy

R. HALABIAN, N. SHAGERDI ESMAEILI, A. OUDI, N. MASROURI, N. AMIRIZADEH, K. MOUSAVI HOSSEINI, A. GHARAH BAGHIAN, H. REZVAN, M.A. JALILI, and M. HABIBI ROUDKENAR, “ISOLATION, CLONING AND EXPRESSION OF RECOMBINANT HUMAN FACTOR VII IN CHO CELL LINE,” THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), vol. 6, no. 1 (22), pp. 1–11, 2009, [Online]. Available: https://sid.ir/paper/78338/en

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