Cost-effective production of tag-less recombinant protein in Nicotiana benthamiana

Md Reyazul Islam, Ju Won Kwak, Jeon soo Lee, Sung Wook Hong, Md Rezaul Islam Khan, Yongjik Lee, Yoontae Lee, Seung Woo Lee, Inhwan Hwang

Research output: Contribution to journalArticlepeer-review


Plants have recently received a great deal of attention as a means of producing recombinant proteins. Despite this, a limited number of recombinant proteins are currently on the market and, if plants are to be more widely used, a cost-effective and efficient purification method is urgently needed. Although affinity tags are convenient tools for protein purification, the presence of a tag on the recombinant protein is undesirable for many applications. A cost-effective method of purification using an affinity tag and the removal of the tag after purification has been developed. The family 3 cellulose-binding domain (CBM3), which binds to microcrystalline cellulose, served as the affinity tag and the small ubiquitin-related modifier (SUMO) and SUMO-specific protease were used to remove it. This method, together with size-exclusion chromatography, enabled purification of human interleukin-6 (hIL6) with a yield of 18.49 mg/kg fresh weight from leaf extracts of Nicotiana benthamiana following Agrobacterium-mediated transient expression. Plant-produced hIL6 (P-hIL6) contained less than 0.2 EU/μg (0.02 ng/mL) endotoxin. P-hIL6 activated the Janus kinase-signal transducer and activator of transcriptional pathways in human LNCaP cells, and induced expression of IL-21 in activated mouse CD4+ T cells. This approach is thus a powerful method for producing recombinant proteins in plants.

Original languageEnglish
Pages (from-to)1094-1105
Number of pages12
JournalPlant Biotechnology Journal
Issue number6
Publication statusPublished - 2019 Jun

Bibliographical note

Funding Information:
This work was supported by the Technology Innovation Program (No. 10063301, Industry core technology development of plant-derived biomaterials for the stem cell culture medium) funded By the Ministry of Trade, Industry & Energy (MOTIE, Korea).

Publisher Copyright:
© 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Agronomy and Crop Science
  • Plant Science


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