PMID: 2488618Jan 1, 1989Paper

Characterisation of a secreted form of recombinant derived human growth hormone, expressed in Escherichia coli cells

Journal of Pharmaceutical and Biomedical Analysis
P GellerforsLinda Fryklund

Abstract

Recombinant DNA derived human growth hormone (rhGH), Genotropin, has been expressed in E. coli cells as a pre-hormone, where the heat stable enterotoxin II signal peptide (STII) was linked to hGH to get secretion of the hormone to the periplasmatic space. The pre-hormone was efficiently cleaved during secretion, by an endogenous signal peptidase generating the correct N-terminal (Phe) end as shown by protein sequence analysis. The purity of rhGH was studied by SDS-PAGE, in combination with laser densitometry and HI-HPLC. These techniques showed that the level of modified rhGH forms, e.g. aggregated and proteolytically cleaved (16 and 6 kDa) in the preparation was in the 0.5-1% range. Furthermore, evidence that the correct disulphide bonds (Cys53-Cys165; Cys182-Cys189) were formed in rhGH during secretion has been shown by a combination of tryptic fingerprint and amino acid analysis. CD-spectroscopic analysis suggested an identical secondary structure to that of pituitary derived human growth hormone (pit-hGH). Isoelectric focusing revealed an isoelectric point (pI) for rhGH of 5.0 similar to pit-hGH and in excellent agreement with the theoretical value 5.1, based on the primary sequence. Finally, an apparent molecular weight of...Continue Reading

References

Nov 1, 1989·Scandinavian Journal of Clinical and Laboratory Investigation·L LannfeltP Gellerfors
Sep 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·S S Abdel-MeguidL A Bentle

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Citations

Jun 1, 1994·Protein Science : a Publication of the Protein Society·E DemchukR C Wade
Nov 22, 1997·Journal of Chromatography. a·S DalmoraP Bartolini
Jun 15, 1992·European Journal of Biochemistry·K AxelssonP Gellerfors
Mar 21, 2009·Biotechnology Progress·Shailaja RabindranVidadi Yusibov
Jan 15, 1994·European Journal of Biochemistry·A M JespersenH H Sørensen
Jun 18, 2003·Journal of Endocrinological Investigation·C L Boguszewski
Jan 1, 1990·Acta Paediatrica Scandinavica. Supplement·P GellerforsS Johansson
Dec 21, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Chulho JungHyun Sik Kim
Nov 10, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jeffrey SchneiderheinzeCharles Demarest

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