Prospects of biosimilar recombinant human growth hormone manufacture in the Republic of Moldova.

Authors

  • Liliana RUSNAC Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”
  • Radu CAZACU Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”
  • Mihail TODIRAȘ Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”
  • Corina SCUTARI Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”
  • Vladimir VALICA Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

DOI:

https://doi.org/10.52692/1857-0011.2022.3-74.30

Keywords:

biosimilar recombinant human growth hormone, domestic pharmaceutical manufacture, technology transfer, Pichia pastoris, pPIC9K plasmid

Abstract

In collaboration of SUMPh „N.Testemițanu”, the pharmaceutical company Balkan Pharmaceuticals and ICGEB the technology transfer during scale-up from the laboratory phase to the pilot phase of the processes of creating the working cell bank and growing Pichia pastoris strains in the Erlenmeyer flask were carried out. Pichia pastoris was selected for recombinant growth hormone as an expression system, which has many advantages over other organisms, including protein folding, post-translational changes in protein, and easy handling. The GH1 altered recombinant human somatropin gene was introduced into the genome of Pichia pastoris culture using restriction near the AOX1 gene promoter as a vector serving the pPIC9K plasmid. The GH1 gene was activated by the AOX1 gene promoter which can be easily induced by the presence of methanol in the culture medium. Subsequently, the „Prepro-alpha Factor Leader” sequence from Saccharomyces cerevisiae was added to the modified sequence so that the obtained somatropin could be secreted into the culture medium. In the technological transfer process the influence of the chosen technological processes on some parameters of the obtained product were examined, Technological Regulations were developed for short and long term storage processes of Pichia pastoris cells and for the process of inoculum preparation, inoculation and growth of P. pastoris strains with recombinant human growth hormone in the Erlenmeyer flask. Standard Operating Procedures were developed for the processes of creating the working cell bank, determining the in-vitro biological activity of the recombinant human growth hormone preparation, with subsequentimplementation in the process of producing rhGH biosimilar at CS Balkan Pharmaceuticals Ltd.

Author Biographies

Liliana RUSNAC, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

dr. în șt. farm., conferențiar cercetător; Centrul Științific al Medicamentului din cadrul

Radu CAZACU, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

Centrul Științific al Medicamentului din cadrul

Mihail TODIRAȘ, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

dr. hab. în șt. med., profesor universitar; Centrul Științific al Medicamentului din cadrul

Corina SCUTARI, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

dr. în șt. med., conferențiar universitar; Catedra de Farmacologie și Farmacie

Vladimir VALICA, Universitatea de Stat de Medicină și Farmacie „Nicolae Testemițanu”

dr. hab. în șt. farm., profesor universitar; Centrul Științific al Medicamentului din cadrul

References

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Published

2022-12-12

Issue

Section

Research Article

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