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Regulated Secretion of Prolactin by the Mouse Insulinoma Cell Line βTC-3

Bio/Technology volume 13, pages 1191–1197 (1995)Cite this article

Abstract

Our aim is to use cultured cells capable of regulated protein secretion for the production of recombi-nant proteins that require particular types of post-translational modifications. Here we have generated a stable transfected βTC-3 cell line, βTC-IPR9, that secretes high levels of recombinant prolactin. Transfected cells synthesize both the 27 kDa glycosylated and a 23 kDa nonglycosylated prolactin; the 23 kDa nonglycosylated species was secreted preferentially when cells were placed in secretion medium containing isobutylmethylxanthine (IBMX) and high concentrations of glucose, K+, and Ca2+. When the cells were cultured in medium containing low concentrations of glucose, K+, and Ca2+, most of the prolactin and insulin were not secreted; much of the prolactin was proteolytically converted to a 16 kDa form. Within the first 30 minutes after transferring the cells to medium containing secretagogues there was a 20-fold increase in the rate of secretion of prolactin; all of the 16 kDa species was secreted. The recombinant cells could be cycled several times between medium in which prolactin was biosynthesized and medium in which it was secreted. Preferential secretion of proteolytically processed prolactin in a medium without contaminating proteins offers an example of the advantage of this technology for production of other recombinant proteins.

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Author information

Author notes

  1. Keqin Chen

    Present address: E.I. Du Pont De Nemours & Company, Central Research & Development, Experimental Station, E328/344A, Wilimington, DE, 19880-0328

Authors and Affiliations

  1. Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Keqin Chen, Gregory N. Stephanopoulos, Anthony J. Sinskey & Harvey F. Lodish

  2. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Gregory N. Stephanopoulos

  3. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Anthony J. Sinskey & Harvey F. Lodish

  4. Whitehead Institute for Biomedical Research, Cambridge, MA, 02142

    Harvey F. Lodish

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  4. Harvey F. Lodish
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Chen, K., Stephanopoulos, G., Sinskey, A. et al. Regulated Secretion of Prolactin by the Mouse Insulinoma Cell Line βTC-3. Nat Biotechnol 13, 1191–1197 (1995). https://doi.org/10.1038/nbt1195-1191

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