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Biological activity and metabolic clearance of recombinant human follicle stimulating hormone produced in Sp2/0 myeloma cells

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Abstract

Human follicle stimulating hormone is a pituitary glycoprotein that is essential for the maintenance of ovarian follicle development and testicular spermatogenesis. Like other members of the glycoprotein hormone family, it contains a common a subunit and a hormone specificβ subunit. Each subunit contains two glycosylation sites. The specific structures of the oligosaccharides of human follicle stimulating hormone have been shown to influence both thein vitro andin vivo bioactivity. Since the carbohydrate structure of a protein reflects the glycosylation apparatus of the host cells in which the protein is expressed, we examined the isoform profiles,in vitro bioactivity and metabolic clearance of a preparation of purified recombinant human follicle stimulating hormone derived from a stable, transfected Sp2/0 myeloma cell line, and pituitary human follicle stimulating hormone. Isoelectric focussing and chromatofocussing studies of human follicle stimulating hormone preparations both showed a more basic isoform profile for the recombinant human follicle stimulating hormone compared to that of pituitary human follicle stimulating hormone. The recombinant human follicle stimulating hormone had a significantly higher radioreceptor activity compared to that of pituitary human follicle stimulating hormone, consistent with a greaterin vitro potency. Pharmacokinetic studies in rats indicated a similar terminal half life (124 min) to that of the pituitary human follicle stimulating hormone (119 min). Preliminary carbohydrate analysis showed recombinant human follicle stimulating hormone to contain high mannose and/or hybrid type, in addition to complex type carbohydrate chains, terminating with bothα2,3 andα2,6 linked sialic acids. These results demonstrate that recombinant human follicle stimulating hormone made in the Sp2/0 myeloma cells is sialylated, has a more basic isoform profile, and has a greaterin vitro biological potency compared to those of the pituitary human follicle stimulating hormone.

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Abbreviations

LH:

Luteinising hormone

FSH:

follicle stimulating hormone

rhFSH:

recombinant human follicle stimulating hormone

TSH:

thyroid stimulating hormone

hCG:

human chorionic gonadotrophin

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Authors and Affiliations

  1. Department of Biotechnology, University of New South Wales, 2033, Kensington, Sydney, Australia

    Catherine K. Chin, Peter P. Gray, Wilaiwan Chotigeat & Stephen M. Mahler

  2. Garvan Institute of Medical Research, St. Vincent's Hospital, 384 Victoria Street, 2010, Darlinghurst, Sydney, Australia

    Peter R. Schofield

  3. Prince Henry's Institute of Medical Research, Monash Medical Centre, 3168, Clayton, Victoria, Australia

    David M. Robertson

Authors
  1. Catherine K. Chin
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  2. Peter R. Schofield
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  3. David M. Robertson
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  4. Peter P. Gray
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  5. Wilaiwan Chotigeat
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  6. Stephen M. Mahler
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Chin, C.K., Schofield, P.R., Robertson, D.M. et al. Biological activity and metabolic clearance of recombinant human follicle stimulating hormone produced in Sp2/0 myeloma cells. Cytotechnology 21, 171–182 (1996). https://doi.org/10.1007/BF02215667

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  • DOI: https://doi.org/10.1007/BF02215667

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