E-ISSN 0976-2779 | ISSN 0975-8453
 

Review Article 


Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha

Susan Zwyea, Rafid A Abdulkareem.

Abstract
A dramatic increase in the amount of recombinant proteins that used in therapeutic applications. TNF-α is one of over 750 recombinant proteins available for use in a broad range of therapeutic applications, including neutrophil chemotaxis, anticoagulatory inhibition, cytolysis and cytostasis of many tumor cell lines. Therefore, in this research Human TNF-α gene has been amplified from the PHA-stimulated peripheral mononuclear blood cells (PBMCs) cDNA pool isolated from healthy volunteers. The amplified human TNF-α gene was combined and was cloned to pPIC9 Plasmid. The fusion gene was expressed in the host strain p.pastoris GS115 using the electroporation technique and inserted under the AOX1promoter's control. Tricine-SDS–PAGE and Western blotting were achieved for methanol-induced expression strain to confirm the successfully secretion of fusion TNF-α protein into the culture medium. Ni-NTA resin was used to purify the recombinant protein. (MTT) assay was used for screening the biological activity of the purified human TNF-α results showed that the Fusion TNF-α reduced the viability of HEP2 cells as long as metabolic activity in dose-dependent manner. Our findings show that TNF-α-gene expressed in the present study is active and had the same pharmacokinetic and biochemical properties as the standard TNF-α and recommended to be used therapeutically as treatment for human diseases, and could be helpful to develop new strategies to cancer therapy.

Key words: hTNF alpha, cloning, Expression, P.Pastoris, biological activity, MTT assay


 
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Pubmed Style

Susan Zwyea, Rafid A Abdulkareem. Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. SRP. 2020; 11(6): 613-621. doi:10.31838/srp.2020.6.91


Web Style

Susan Zwyea, Rafid A Abdulkareem. Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. http://www.sysrevpharm.org//?mno=117088 [Access: June 21, 2020]. doi:10.31838/srp.2020.6.91


AMA (American Medical Association) Style

Susan Zwyea, Rafid A Abdulkareem. Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. SRP. 2020; 11(6): 613-621. doi:10.31838/srp.2020.6.91



Vancouver/ICMJE Style

Susan Zwyea, Rafid A Abdulkareem. Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. SRP. (2020), [cited June 21, 2020]; 11(6): 613-621. doi:10.31838/srp.2020.6.91



Harvard Style

Susan Zwyea, Rafid A Abdulkareem (2020) Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. SRP, 11 (6), 613-621. doi:10.31838/srp.2020.6.91



Turabian Style

Susan Zwyea, Rafid A Abdulkareem. 2020. Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. Systematic Reviews in Pharmacy, 11 (6), 613-621. doi:10.31838/srp.2020.6.91



Chicago Style

Susan Zwyea, Rafid A Abdulkareem. "Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha." Systematic Reviews in Pharmacy 11 (2020), 613-621. doi:10.31838/srp.2020.6.91



MLA (The Modern Language Association) Style

Susan Zwyea, Rafid A Abdulkareem. "Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha." Systematic Reviews in Pharmacy 11.6 (2020), 613-621. Print. doi:10.31838/srp.2020.6.91



APA (American Psychological Association) Style

Susan Zwyea, Rafid A Abdulkareem (2020) Cloning, Expression and Bioactivity of Human Tumor Necrosis Factor Alpha. Systematic Reviews in Pharmacy, 11 (6), 613-621. doi:10.31838/srp.2020.6.91





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