Expression and secretion of a biologically active glycoprotein hormone, ovine follicle stimulating hormone, by Pichia Pastoris

Citation
Ae. Fidler et al., Expression and secretion of a biologically active glycoprotein hormone, ovine follicle stimulating hormone, by Pichia Pastoris, J MOL ENDOC, 21(3), 1998, pp. 327-336
Citations number
34
Categorie Soggetti
Endocrinology, Nutrition & Metabolism
Journal title
JOURNAL OF MOLECULAR ENDOCRINOLOGY
ISSN journal
09525041 → ACNP
Volume
21
Issue
3
Year of publication
1998
Pages
327 - 336
Database
ISI
SICI code
0952-5041(199812)21:3<327:EASOAB>2.0.ZU;2-E
Abstract
The methylotrophic yeast, Pichia pastoris, has been used to co-express reco mbinant genes formed by fusion of the mating factor-alpha (MF alpha) leader and ovine follicle stimulating hormone (oFSH) alpha and beta subunit codin g sequences. Pichia strains carrying single copies of the two fusion genes secreted recombinant oFSH (roFSH) to concentrations of approximately 51.0 n g/ml and 17.5 ng/ml, measured by RIA or in vitro bioassay respectively, whe reas a strain with two copies of the alpha and one copy of the beta subunit fusion genes secreted roFSH to concentrations of 61 ng/ml (RIA) and 22 ng/ ml (bioassay). It appears that the Pichia-derived roFSH had about one-third the in vitro bioactivity of native oFSH or, alternatively, only one-third of the roFSH is bioactive. Measurements of secreted roFSH alpha and beta su bunit concentrations indicated less than 10% of alpha and 25-33% af beta su bunits were stably dimerized. The receptor binding properties of the roFSH resemble those of native oFSH. In summary this paper reports the production , by P. pastoris, of a heterodimeric glycoprotein hormone (roFSH) that has in vitro biological activity.