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. 1966 Sep;100(3):711–717. doi: 10.1042/bj1000711

Isolation and properties of the electrophoretic components of human growth hormone by Sephadex-gel filtration and preparative polyacrylamide-gel electrophoresis

B B Saxena 1,*, P H Henneman 1
PMCID: PMC1265205  PMID: 16742415

Abstract

1. Human growth hormone was prepared from acetone-dried residues after extraction of gonadotrophins from pituitary glands. 2. Crude growth hormone was purified by gel filtration on Sephadex, resulting in a product that is soluble in water or 0·5% sodium chloride. It is painless on injection and shows a twofold increase in biological potency. Aggregation of growth hormone on Sephadex columns can be avoided by the addition of urea (6m) and EDTA (1mm) to the buffer. 3. Growth hormone appeared as a single component from Sephadex and ion-exchange columns and sedimented as a single boundary in the ultracentrifuge. In the circular disk electrophoresis, however, the growth hormone showed one faster and two slower-moving anionic components. 4. These components were isolated by preparative electrophoresis on polyacrylamide columns. The purified growth hormone and its three components sedimented as single boundaries with coefficients 2·62, 2·66, 2·66 and 2·83s respectively. 5. Amino acid analyses of the purified growth hormone and its components were closely related. End-group analysis of purified growth hormone and its components showed only phenylalanine at both N- and C-terminals. 6. The purified growth hormone and its components were essentially free of other pituitary hormones, but contained significant prolactin activity. The biochemical similarities among the electrophoretic components of human growth hormone and the presence of the same three components in the growth hormone prepared from a single human pituitary gland suggest polymorphism of a biologically active protein molecule.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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