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Production of Recombinant Human Growth Hormone Conjugated with a Transcytotic Peptide in Pichia pastoris for Effective Oral Protein Delivery

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Abstract

Among the possible delivery routes, the oral administration of a protein is simple and achieves high patient compliance without pain. However, the low bioavailability of a protein drug in the intestine due to the physical barriers of the intestinal epithelia is the most critical problem that needs to be solved. To overcome the low bioavailability of a protein drug in the intestine, we aimed to construct a recombinant Pichia pastoris expressing a human growth hormone (hGH) fusion protein conjugated with a transcytotic peptide (TP) that was screened through peroral phage display to target goblet cells in the intestinal epithelia. The TP-conjugated hGH was successfully produced in P. pastoris in a secreted form at concentrations of up to 0.79 g/l. The function of the TP-conjugated hGH was validated by in vitro and in vivo assays. The transcytotic function of the TP through the intestinal epithelia was verified only in the C terminus conjugated hGH, which demonstrated the induction of IGF-1 in a HepG2 cell culture assay, a higher translocation of recombinant hGH into the ileal villi after oral administration in rats and both IGF-1 induction and higher body weight gain in rats after oral administration. The present study introduces the possibility for the development of an effective oral protein delivery system in the pharmaceutical and animal industries through the introduction of an effective TP into hGH.

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Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2011-0029416). We also acknowledge the National Instrumental Centre for Environmental Management (NICEM) and National Center for Inter-University Research Facilities (NCIRF). Jun-Yeong Lee was supported by BK21 program.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Republic of Korea

    Jun-Yeong Lee, Hui-Shan Li, Chang-Yun Choi, Tae-Eun Park & Yun-Jaie Choi

  2. Institute of Green-Bio Science & Technology, Seoul National University, Pyeongchanggun, Gangwondo, 232-916, Republic of Korea

    Sang-Kee Kang & Jin-Duck Bok

  3. Department of Nano-Bioengineering, College of Life Science and Bioengineering, University of Incheon, Incheon, 406-772, Republic of Korea

    Seung-Ho Lee

  4. Research Institute for Agriculture and Life Science, Seoul National University, Seoul, 151-921, Republic of Korea

    Chong-Su Cho & Yun-Jaie Choi

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  1. Jun-Yeong Lee
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  2. Sang-Kee Kang
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  3. Hui-Shan Li
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  7. Seung-Ho Lee
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  9. Yun-Jaie Choi
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Correspondence to Jun-Yeong Lee or Yun-Jaie Choi.

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Jun-Yeong Lee and Sang-Kee Kang have contributed equally to this work.

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Lee, JY., Kang, SK., Li, HS. et al. Production of Recombinant Human Growth Hormone Conjugated with a Transcytotic Peptide in Pichia pastoris for Effective Oral Protein Delivery. Mol Biotechnol 57, 430–438 (2015). https://doi.org/10.1007/s12033-014-9835-0

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