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High-Level Expression, Purification and Production of the Fungal Immunomodulatory Protein-Gts in Baculovirus-Infected Insect Larva

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Abstract

Fip-gts, a fungal immunomodulatory protein (Fip) isolated from Ganoderma tsugae (gts), has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. To cost-effectively produce Fip-gts and bypass the bottleneck involved in its time-consuming purification from G. tsugae, in this study, we incorporated the SPbbx secretion signal into recombinant baculovirus for expressing glycosylated and bioactive rFip-gts in baculovirus-infected insect cells and Trichoplusia ni larva. This is the first study to employ the aerosol infecting T. ni larva with recombinant baculovirus for economical and high-level production of foreign proteins. In this study, one purification could yield 10 mg of rFip-gts protein merely from ∼100 infected T. ni larvae by aerosol inoculation, corresponding to 5 L (5 × 109 cells) of the infected Sf21 culture. In addition, the rFip-gts purified from T. ni larvae could induce the expression of interleukin-2 in murine splenocytes with an immunoresponsive level similar to that induced by LZ-8 (a known potent immunomodulatory protein purified from Ling zhi, Ganoderma lucidum). Thus, our results demonstrated that the larva-based baculovirus expression system can successfully express rFip-gts with the assembling capability required for maintaining immunomodulatory and anticancer activity. Our approach will open a new avenue for the production of rFip-gts and facilitate the immunoregulatory activity of rFip-gts available in the future.

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Acknowledgments

The authors would like to thank the China Medical University and National Science Council of the Republic of China for financially supporting this research under Contract No. CMU 97-199 and NSC-98-2324-B-039-005.

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Authors and Affiliations

  1. Department of Science Technology, Chung Yuan Christian University, Chung Li, Taiwan, 32023, Republic of China

    Tzong-Yuan Wu

  2. Division of General Surgery, Department of Surgery, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, 11031, Republic of China

    Hsin-An Chen

  3. Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan, 11031, Republic of China

    Hsin-An Chen

  4. Department of Chemistry, National Chung Hsing University, Taichung, Taiwan, 40201, Republic of China

    Feng-Yin Li

  5. School of Chinese Medicine, China Medical University, Taichung, Taiwan, 40402, Republic of China

    Ching-Ting Lin & Tzyy-Rong Jinn

  6. Graduate Institute of Veterinary Pathobiology, China Medical University, Taichung, Taiwan, 40402, Republic of China

    Chi-Ming Wu

  7. Biopesticide Department, Agricultural Chemicals and Toxic Substances Research Institute, Taichung, Taiwan, 41300, Republic of China

    Feng-Chia Hsieh

  8. Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan, 40201, Republic of China

    Jason Tze-Cheng Tzen & Sheng-Kuo Hsieh

  9. Institute of Medical and Molecular Toxicology, Chung Shan Medical University, Taichung, Taiwan, 40201, Republic of China

    Jiunn-Liang Ko

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  1. Tzong-Yuan Wu
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Correspondence to Tzyy-Rong Jinn.

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Tzong-Yuan Wu and Hsin-An Chen contributed equally to this study.

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Wu, TY., Chen, HA., Li, FY. et al. High-Level Expression, Purification and Production of the Fungal Immunomodulatory Protein-Gts in Baculovirus-Infected Insect Larva. Appl Biochem Biotechnol 169, 976–989 (2013). https://doi.org/10.1007/s12010-012-0049-2

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