title: Production of recombinant human growth hormone conjugated with a transcytotic peptide in Pichia pastoris for effective oral protein delivery.
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.
authors.affiliation: Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Republic of Korea, akirus86@snu.ac.kr.
authors.forename: Jun-Yeong
authors.initials: JY
authors.lastname: Lee
authors.forename: Sang-Kee
authors.initials: SK
authors.lastname: Kang
authors.forename: Hui-Shan
authors.initials: HS
authors.lastname: Li
authors.forename: Chang-Yun
authors.initials: CY
authors.lastname: Choi
authors.forename: Tae-Eun
authors.initials: TE
authors.lastname: Park
authors.forename: Jin-Duck
authors.initials: JD
authors.lastname: Bok
authors.forename: Seung-Ho
authors.initials: SH
authors.lastname: Lee
authors.forename: Chong-Su
authors.initials: CS
authors.lastname: Cho
authors.forename: Yun-Jaie
authors.initials: YJ
authors.lastname: Choi
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002463:Cell Membrane Permeability; D003001:Cloning, Molecular; D056945:Hep G2 Cells; D019382:Human Growth Hormone / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008297:Male; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000378:metabolism; D057900:Transcytosis
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D019382:Human Growth Hormone; D007334:Insulin-Like Growth Factor I