High-level expression, purification and production of the fungal immunomodulatory protein-gts in baculovirus-infected insect larva.

Tzong-Yuan Wu, Hsin-An Chen, Feng-Yin Li, Ching-Ting Lin, Chi-Ming Wu, Feng-Chia Hsieh, Jason Tze-Cheng Tzen, Sheng-Kuo Hsieh, Jiunn-Liang Ko, Tzyy-Rong Jinn

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

Applied biochemistry and biotechnology 2013 Feb

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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 SP(bbx) 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.

Citation

Tzong-Yuan Wu, Hsin-An Chen, Feng-Yin Li, Ching-Ting Lin, Chi-Ming Wu, Feng-Chia Hsieh, Jason Tze-Cheng Tzen, Sheng-Kuo Hsieh, Jiunn-Liang Ko, Tzyy-Rong Jinn. High-level expression, purification and production of the fungal immunomodulatory protein-gts in baculovirus-infected insect larva. Applied biochemistry and biotechnology. 2013 Feb;169(3):976-89