Practical reversed-phase high-performance liquid chromatography method for laboratory-scale purification of recombinant human thyrotropin

Journal of Chromatography. a
João Ezequiel OliveiraMaria Teresa Carvalho Pinto Ribela


A small, semi-preparative C(4) RP-HPLC column was used to set up the conclusive laboratory-scale purification of Chinese hamster ovary-derived human thyrotropin (hTSH), after a preliminary concentration-purification of an extremely dilute and poorly ( approximately 0.6 microg hTSH/mL; mass fraction=0.35%) conditioned medium on a cation exchanger. Several fractions of this eluate were repeatedly injected on the semi-preparative column, obtaining, in a single run (<1h chromatographic time), a concentrated pool ( approximately 1.2 mg/mL) of highly purified hTSH that could be further concentrated to >3 mg/mL and then efficiently lyophilized. The overall recovery in the rapid RP-HPLC purification step, including concentration and lyophilization, was of the order of 80%. The final product, when tested via a precise, single-dose in vivo bioassay, confirmed that it did not suffer any loss of bioactivity. This same methodology can be easily adapted to the small-scale purification of other recombinant products, even when obtained from genetically modified organisms at extremely low concentrations and mass fractions.


Aug 4, 1989·Journal of Chromatography·R BischoffC Roitsch
Jul 27, 1984·Journal of Chromatography·D L UrdalS Gillis
Feb 1, 1995·Thyroid : Official Journal of the American Thyroid Association·J East-PalmerB D Weintraub
Nov 22, 1997·Journal of Chromatography. a·S DalmoraP Bartolini
Jul 26, 2000·Journal of Chromatography. a·S RamananA Velayudhan
Jun 8, 2001·Biotechnology and Applied Biochemistry·A BeldarraínM Gil
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·João Ezequiel de OliveiraMaria Teresa C P Ribela
May 28, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Maria Teresa C P RibelaPaolo Bartolini
Oct 20, 2006·Journal of Chromatography. a·Renan Fernandes LoureiroMaria Teresa C P Ribela

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Jun 14, 2012·Applied Biochemistry and Biotechnology·Fernanda Santos ArthusoCarlos Roberto Jorge Soares
Sep 3, 2013·Applied Biochemistry and Biotechnology·Renata DamianiMaria Teresa C P Ribela
Jan 27, 2009·Journal of Chromatography. a·C M CarvalhoM T C P Ribela

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