TRANSIENT AND STABLE EXPRESSION OF BIOLOGICALLY ACTIVE RECOMBINANT B-DOMAIN-DELETED HUMAN FACTOR VIII IN MAMMALIAN CELLS

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

AMIRI ZAHDEH N.; POURFATH ELAH A.A.; ZOMORODIPOUR A.; ATAEI FARNAZ; DIZAJI A.A.KH.; LENJAN NEZHADIAN H.; KHODABANDEH M.; RASTGAR LARI GH.; SALEK M.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: Journal of Sciences, Islamic Republic of Iran Year:2005 | Volume:16 | Issue:2 Page(s): 103-112

Download Full-Text

View:

417

Download:

159

Cites:

Information Journal Paper

Title

TRANSIENT AND STABLE EXPRESSION OF BIOLOGICALLY ACTIVE RECOMBINANT B-DOMAIN-DELETED HUMAN FACTOR VIII IN MAMMALIAN CELLS

Author(s)

Keywords

RECOMBINANT HUMAN COAGULATION FACTOR VIII; MAMMALIAN CELL EXPRESSION SYSTEM; CHO AND COS7; HEMOPHILIA A

Abstract

Hemophilia A is an X-linked recessive bleeding disorder, widely prevalent throughout the world, for which, replacement therapy is a current treatment done by infusion of either human plasma derived FVIII or recombinant FVIII. In order to produce a recombinant form of biologically active human coagulation factor VIII, a mammalian expression system is necessary for proper post-translational modifications. In this regard, two types of mammalian cell lines, COS7 and CHO, were transfected with a recombinant plasmid, constructed by insertion of a NotI restriction fragment containing B-domain-deleted cDNA of hFVIII in pcDNA3 plasmid, downstream of CMV promoter. By performing one-stage clotting assay as well as ELISA test on the conditioned media collected from transfected cells, we confirmed transient and stable expression of rhFVIII in the transfected COS7 and CHO cells, respectively. The presence of rhFVIII mRNA was also demonstrated by performing RT-PCR on total cellular RNA, extracted from the stably transfected CHO cells. The highest amount of produced active rhFVIII in the stably transfected CHO cells was estimated to be around 0.1 U/ml of the cultured media. By applying southern blotting experiment on the digested as well as high molecular weight chromosomal DNA, prepared from the stably transfected CHO cells, we have demonstrated the presence of the rhFVIII expressing plasmid in the CHO cells. The recombinant plasmid as well as the stable FVIII expressing cell line developed in this study has provided useful bases for further molecular studies of various important factors influencing the expression efficiency of rhFVIII.

Cites

BACTERIAL EXPRESSION AND PURIFICATION OF C1C2 DOMAIN OF HUMAN FACTOR VIII

References

No record.

Cite

APA: Copy

AMIRI ZAHDEH, N., POURFATH ELAH, A.A., ZOMORODIPOUR, A., ATAEI, FARNAZ, DIZAJI, A.A.KH., LENJAN NEZHADIAN, H., KHODABANDEH, M., RASTGAR LARI, GH., & SALEK, M.. (2005). TRANSIENT AND STABLE EXPRESSION OF BIOLOGICALLY ACTIVE RECOMBINANT B-DOMAIN-DELETED HUMAN FACTOR VIII IN MAMMALIAN CELLS. JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN, 16(2), 103-112. SID. https://sid.ir/paper/529617/en

Vancouver: Copy

AMIRI ZAHDEH N., POURFATH ELAH A.A., ZOMORODIPOUR A., ATAEI FARNAZ, DIZAJI A.A.KH., LENJAN NEZHADIAN H., KHODABANDEH M., RASTGAR LARI GH., SALEK M.. TRANSIENT AND STABLE EXPRESSION OF BIOLOGICALLY ACTIVE RECOMBINANT B-DOMAIN-DELETED HUMAN FACTOR VIII IN MAMMALIAN CELLS. JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN[Internet]. 2005;16(2):103-112. Available from: https://sid.ir/paper/529617/en

IEEE: Copy

N. AMIRI ZAHDEH, A.A. POURFATH ELAH, A. ZOMORODIPOUR, FARNAZ ATAEI, A.A.KH. DIZAJI, H. LENJAN NEZHADIAN, M. KHODABANDEH, GH. RASTGAR LARI, and M. SALEK, “TRANSIENT AND STABLE EXPRESSION OF BIOLOGICALLY ACTIVE RECOMBINANT B-DOMAIN-DELETED HUMAN FACTOR VIII IN MAMMALIAN CELLS,” JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN, vol. 16, no. 2, pp. 103–112, 2005, [Online]. Available: https://sid.ir/paper/529617/en

Related Journal Papers

No record.

Related Seminar Papers

No record.

Related Plans

No record.

Recommended Workshops