Structural and functional characterization of recombinant human growth hormone isolated from transgenic pig milk

PLoS One. 2020 Jul 31;15(7):e0236788. doi: 10.1371/journal.pone.0236788. eCollection 2020.

Abstract

This study aimed to establish and reproduce transgenic pigs expressing human growth hormone (hGH) in their milk. We also aimed to purify hGH from the milk, to characterize the purified protein, and to assess the potential of our model for mass production of therapeutic proteins using transgenic techniques. Using ~15.5 L transgenic pig milk, we obtained proteins with ≥ 99% purity after three pre-treatments and five column chromatography steps. To confirm the biosimilarity of our milk-derived purified recombinant hGH (CGH942) with commercially available somatropin (Genotropin), we performed spectroscopy, structural, and biological analyses. We observed no difference between the purified protein and Genotropin samples. Furthermore, rat models were used to assess growth promotion potential. Our results indicate that CGH942 promotes growth, by increasing bone development and body weight. Toxicity assessments revealed no abnormal findings after 4 weeks of continuous administration and 2 weeks of recovery. The no-observed-adverse-effect level for both males and females was determined to be 0.6 mg/kg/day. Thus, no toxicological differences were observed between commercially available somatropin and CGH942 obtained from transgenic pig milk. In conclusion, we describe a transgenic technique using pigs, providing a new platform to produce human therapeutic proteins.

MeSH terms

  • Animals
  • Animals, Genetically Modified / metabolism*
  • Chromatography, Affinity
  • Female
  • Gene Transfer Techniques
  • Human Growth Hormone* / chemistry
  • Human Growth Hormone* / genetics
  • Human Growth Hormone* / metabolism
  • Humans
  • Rats
  • Recombinant Proteins* / chemistry
  • Recombinant Proteins* / metabolism
  • Recombinant Proteins* / toxicity
  • Swine

Substances

  • Recombinant Proteins
  • Human Growth Hormone

Grants and funding

Many authors are employed by CHO-A, which is a pharmaceutical company. The company (the funder) provided support in the form of salaries for authors but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.