Literature DB >> 18295789

A novel reporter gene assay for interferons based on CHO-K1 cells.

V Smilović1, S Caserman, I Fonda, V Gaberc-Porekar, V Menart.   

Abstract

Interferons (IFNs) are cytokines playing an important role in the immune response and defence against viruses. They are widely used as biopharmaceuticals. Currently, the anti-viral assay (AVA) is the most commonly used bioassay for determining interferon potency. In the search for rapid and robust but reliable methods, reporter gene assays (RGA) appear to be the most promising approach, therefore we have designed a new reporter cell line, CHO-ISRE-SEAP, suitable for determination of type I interferon potency. Chinese hamster ovary (CHO-K1) cells were stably transfected with secretory alkaline phosphatase (SEAP) gene under the control of interferon stimulated response element (ISRE) promoter. The amount of SEAP in the cell culture medium can be easily measured colorimetrically and has been found to correlate with the amount of IFN added. The new assay is widely applicable for determination of type I IFNs, such as IFN-alpha, IFN-beta and IFN-omega, in research, development of IFN biopharmaceuticals, in batch release, etc. Interestingly, in this assay, IFN-beta shows approximately 6 times higher response than IFN-alpha, which makes it especially appropriate for measuring low levels of IFN-beta. Compared to other known RGAs, the novel CHO-ISRE-SEAP cell line-based RGA appears to have certain advantages with respect to cost and performance.

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Year:  2008        PMID: 18295789     DOI: 10.1016/j.jim.2008.01.011

Source DB:  PubMed          Journal:  J Immunol Methods        ISSN: 0022-1759            Impact factor:   2.303


  4 in total

1.  Induction of IFN-α subtypes and their antiviral activity in mumps virus infection.

Authors:  Maja Markušić; Maja Šantak; Tanja Košutić-Gulija; Mladen Jergović; Renata Jug; Dubravko Forčić
Journal:  Viral Immunol       Date:  2014-12       Impact factor: 2.257

2.  In vitro evaluation of antioxidant and radioprotective properties of a novel extremophile from mud volcano: implications for management of radiation emergencies.

Authors:  Atlar Singh Dhaker; Rohit Marwah; Rakesh Damodar; Damodar Gupta; Hemant Kumar Gautam; Sarwat Sultana; Rajesh Arora
Journal:  Mol Cell Biochem       Date:  2011-04-03       Impact factor: 3.396

3.  Bioactivity determination of native and variant forms of therapeutic interferons.

Authors:  Louise Larocque; Alex Bliu; Ranran Xu; Abebaw Diress; Junzhi Wang; Rongtuan Lin; Runtao He; Michel Girard; Xuguang Li
Journal:  J Biomed Biotechnol       Date:  2011-03-03

4.  Validation of a cell-based colorimetric reporter gene assay for the evaluation of Type I Interferons.

Authors:  Ignacio Mejía-Calvo; Leslie Muñoz-García; Alexis Jiménez-Uribe; Rosa Camacho-Sandoval; Edith González-González; Gabriela Mellado-Sánchez; Alejandra V Tenorio-Calvo; Carlos A López-Morales; Marco A Velasco-Velázquez; Lenin Pavón; Sonia Mayra Pérez-Tapia; Emilio Medina-Rivero
Journal:  Biotechnol Rep (Amst)       Date:  2019-04-18
  4 in total

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