Repository of Research and Investigative Information

Repository of Research and Investigative Information

Rafsanjan University of Medical Sciences

Generation of truncated recombinant form of tumor necrosis factor receptor-1 to produce cancer vaccine

(2016) Generation of truncated recombinant form of tumor necrosis factor receptor-1 to produce cancer vaccine. Tropical Journal of Pharmaceutical Research. pp. 2345-2350. ISSN 1596-5996

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Abstract

Purpose: To produce truncated recombinant form of tumor necrosis factor receptor 1 (TNFR1), cysteine-rich domain 2 (CRD2) and CRD3 regions of the receptor were generated using pET28a and E. coli/BL21. Methods: DNA coding sequence of CRD2 and CRD3 was cloned into pET28a vector and the corresponding protein was expressed under induction of isopropyl beta-D-1-thiogalactopyranoside (IPTG) as 6xHis tagged using E. coli BL21 (DE3) expression system. The protein was then purified by Ni-NTA affinity chromatography. The fragment insertion, expression of recombinant protein and the yield of expression were evaluated. Results: Protein expression was achieved by identifying a band with molecular weight of 1488.3 Da. The recombinant protein of CRD2 and CRD3 was most efficiently expressed in 0.5 mM IPTG and 3 h of incubation at 37 degrees C with high yield equal to 0.3 mu g/mu l. Also, the highest concentration of imidazole for purification of the recombinant protein was 250 mM. Conclusion: A truncated form of TNFR-1 has been successfully expressed in a bacterial expression system and purified on affinity column. The purified protein can be used in in vivo experiments to prepare specified agonist antibodies for TNFR-1.

Item Type: Article
Keywords: Tumor necrosis factor receptor 1 (TNFR-1) Cysteine rich domain 2 (CRD2) Cysteine rich domain (CRD3) Apoptosis Cancer vaccine Antibodies Recombinant protein pET28a nf-kappa-b tnf receptor therapeutic strategy activation alpha complex Pharmacology & Pharmacy
Page Range: pp. 2345-2350
Journal or Publication Title: Tropical Journal of Pharmaceutical Research
Journal Index: ISI
Volume: 15
Number: 11
Identification Number: https://doi.org/10.4314/tjpr.v15i11.7
ISSN: 1596-5996
Depositing User: مهندس مهدی شریفی
URI: http://eprints.rums.ac.ir/id/eprint/4448

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