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

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

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)976-989
Number of pages14
JournalApplied Biochemistry and Biotechnology
Volume169
Issue number3
DOIs
Publication statusPublished - Feb 2013

Fingerprint

Ganoderma
Fungal Proteins
Baculoviridae
Purification
Larva
Insects
Aerosols
Proteins
Interleukin-2
Reishi
Costs
Sf9 Cells
Therapeutic Uses
Autoimmune Diseases

Keywords

  • Aerosol infection
  • Baculovirus expression system
  • Fungal immunomodulatory protein
  • Glycoprotein
  • Ling zhi
  • Trichoplusia ni larva

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Environmental Engineering

Cite this

High-level expression, purification and production of the fungal immunomodulatory protein-gts in baculovirus-infected insect larva. / Wu, Tzong Yuan; Chen, Hsin-An; Li, Feng Y.; Lin, Ching Ting; Wu, Chi Ming; Hsieh, Feng Chia; Tzen, Jason Tze Cheng; Hsieh, Sheng Kuo; Ko, Jiunn Liang; Jinn, Tzyy Rong.

In: Applied Biochemistry and Biotechnology, Vol. 169, No. 3, 02.2013, p. 976-989.

Research output: Contribution to journalArticle

Wu, Tzong Yuan ; Chen, Hsin-An ; Li, Feng Y. ; Lin, Ching Ting ; Wu, Chi Ming ; Hsieh, Feng Chia ; Tzen, Jason Tze Cheng ; Hsieh, Sheng Kuo ; Ko, Jiunn Liang ; Jinn, Tzyy Rong. / High-level expression, purification and production of the fungal immunomodulatory protein-gts in baculovirus-infected insect larva. In: Applied Biochemistry and Biotechnology. 2013 ; Vol. 169, No. 3. pp. 976-989.
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AU - Lin, Ching Ting

AU - Wu, Chi Ming

AU - Hsieh, Feng Chia

AU - Tzen, Jason Tze Cheng

AU - Hsieh, Sheng Kuo

AU - Ko, Jiunn Liang

AU - Jinn, Tzyy Rong

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AB - 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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