AFFINITY PURIFICATION OF TUMOR NECROSIS FACTOR-α EXPRESSED IN RAJI CELLS BY PRODUCED SCFV ANTIBODY COUPLED CNBR-ACTIVATED SEPHAROSE

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ABD ALALI ZADEH JALAL; MAJIDI ZOLBANIN JAFAR; NOURI MOHAMMAD; BARADARAN BEHZAD; MOVASSAGHPUOR ALI AKBAR; FARAJNIA SAFAR; OMIDI YADOLLAH

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Journal: Advanced Pharmaceutical Bulletin Year:2013 | Volume:3 | Issue:1 Page(s): 19-23

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Title

AFFINITY PURIFICATION OF TUMOR NECROSIS FACTOR-α EXPRESSED IN RAJI CELLS BY PRODUCED SCFV ANTIBODY COUPLED CNBR-ACTIVATED SEPHAROSE

Author(s)

Keywords

TNF-α EXPRESSION

AFFINITY PURIFICATION

MONOCLONAL ANTIBODY

LPS

Abstract

Purpose: Recombinant tumor necrosis factor-alpha (TNF-a) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins.Methods: In this study, we examined the potential of our produced anti-TNF-a scFv fragments for purification of TNF-a produced by Raji cells. The Raji cells were induced by lipopolysaccharides (LPS) to express TNF-a Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-a expression. The anti-TNF-a scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for AFFINITY PURIFICATION of expressed TNF-a and the purity of the protein was assessed by SDS-PAGE.Results: Western blot and FACS flow cytometry analyses showed the successful expression of TNF-a with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-a protein with purity over 95%.Conclusion: These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-a protein can be applied for various in vitro and in vivo applications.

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APA: Copy

ABD ALALI ZADEH, JALAL, MAJIDI ZOLBANIN, JAFAR, NOURI, MOHAMMAD, BARADARAN, BEHZAD, MOVASSAGHPUOR, ALI AKBAR, FARAJNIA, SAFAR, & OMIDI, YADOLLAH. (2013). AFFINITY PURIFICATION OF TUMOR NECROSIS FACTOR-α EXPRESSED IN RAJI CELLS BY PRODUCED SCFV ANTIBODY COUPLED CNBR-ACTIVATED SEPHAROSE. ADVANCED PHARMACEUTICAL BULLETIN, 3(1), 19-23. SID. https://sid.ir/paper/331701/en

Vancouver: Copy

ABD ALALI ZADEH JALAL, MAJIDI ZOLBANIN JAFAR, NOURI MOHAMMAD, BARADARAN BEHZAD, MOVASSAGHPUOR ALI AKBAR, FARAJNIA SAFAR, OMIDI YADOLLAH. AFFINITY PURIFICATION OF TUMOR NECROSIS FACTOR-α EXPRESSED IN RAJI CELLS BY PRODUCED SCFV ANTIBODY COUPLED CNBR-ACTIVATED SEPHAROSE. ADVANCED PHARMACEUTICAL BULLETIN[Internet]. 2013;3(1):19-23. Available from: https://sid.ir/paper/331701/en

IEEE: Copy

JALAL ABD ALALI ZADEH, JAFAR MAJIDI ZOLBANIN, MOHAMMAD NOURI, BEHZAD BARADARAN, ALI AKBAR MOVASSAGHPUOR, SAFAR FARAJNIA, and YADOLLAH OMIDI, “AFFINITY PURIFICATION OF TUMOR NECROSIS FACTOR-α EXPRESSED IN RAJI CELLS BY PRODUCED SCFV ANTIBODY COUPLED CNBR-ACTIVATED SEPHAROSE,” ADVANCED PHARMACEUTICAL BULLETIN, vol. 3, no. 1, pp. 19–23, 2013, [Online]. Available: https://sid.ir/paper/331701/en

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