Enhanced production and single step purification of biologically active recombinant anti-IL6 scFv from Escherichia coli inclusion bodies

Şahinbaş D., Çelik E.

Process Biochemistry, vol.133, pp.151-157, 2023 (SCI-Expanded) identifier

  • Publication Type: Article / Article
  • Volume: 133
  • Publication Date: 2023
  • Doi Number: 10.1016/j.procbio.2023.08.013
  • Journal Name: Process Biochemistry
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aerospace Database, Aqualine, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Communication Abstracts, Compendex, Food Science & Technology Abstracts, INSPEC, Metadex, Pollution Abstracts, Veterinary Science Database, Civil Engineering Abstracts
  • Page Numbers: pp.151-157
  • Keywords: Escherichia coli, Inclusion bodies, Interleukin-6, N-lauroyl sarcosine, Single chain fragment variable (scFv)
  • Hacettepe University Affiliated: Yes

Abstract

Interleukin 6 (IL-6) is a proinflammatory cytokine that plays a critical role in immune responses and inflammation. Overexpression of IL-6 serves as a biomarker for various diseases. Therefore, it is vital to develop IL-6-specific antibodies for the diagnosis and treatment of inflammatory diseases. Alternatively, high-yield production of single-chain variable fragments (scFv) in prokaryotes reduces the time and cost required while attaining a similar specificity. However, overexpression of eukaryotic proteins in bacterial expression systems often leads to the formation of inclusion bodies. Here, a codon-optimized anti-IL6 scFv was expressed in the E. coli SHuffle strain, and the effects of inclusion body solubilisation methods on the product yield were investigated. A high concentration (23 mg/L) of biologically active and pure (>95 %) scFv was obtained without any need for a refolding step following solubilisation with a non-denaturing solubilisation agent, N-lauroyl sarcosine. Activity assays showed that anti-IL6 scFv could specifically bind to its antigen. Overall, we present reproducible and efficient production of anti-IL6 scFv in E. coli by using a one-step purification method. While the biologically active and pure antibody fragment can be used for diagnostic studies of the IL-6 cytokine, the developed method can be applied to other recombinant proteins expressed in E. coli.