Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents.

Samira Nekoufar,Mohammad Reza Fazeli,Ahmad Fazeli

doi:10.34172/apb.2020.027

Samira Nekoufar, Mohammad Reza Fazeli + Show 1 more

Open Access

https://doi.org/10.34172/apb.2020.027

Copy DOI

Abstract

Purposes: Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon β-1b inclusion bodies. Methods: Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon β-1b inclusion bodies. Results: Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon β-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% (v/v) 1-propanol and 20% (v/v) 2-butanol into the buffer solution as well. Conclusion: The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area.

Highlights

Multiple sclerosis (MS) is an inflammatory chronic disease characterized by multiple neurological symptoms and disability which involves the nervous system.[1]
We have demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3
The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area

Summary

Introduction

Multiple sclerosis (MS) is an inflammatory chronic disease characterized by multiple neurological symptoms and disability which involves the nervous system.[1]. Environment, and infection (e.g. viruses) may be the factors that are involved in the MS pathogenesis.[1] So far, interferon B is a drug of choice to alleviate relapsing forms of MS which was discovered for the first time by Issacs and Lindenmann in 1957.2,3 This medication is administered either as INFβ-1a; a natural form[1] or INFβ1b; a non-glycosylated form.[4,5,6] The amount which the human body produces is not sufficient to fully cure the disease, finding another source is necessary.

Objectives

Methods

Results

Conclusion