| Title | Isolation and purification of thyroid-stimulating hormone from human pituitaries by affinity chromatography | |
| Author | Helen Ath. Dimitriou
Medical School, University of Crete, 713 52, Iraklion, Crete, Greece |
|
| Citation | Dimitriou, H.A.: Isolation and purification of thyroid-stimulating hormone from human pituitaries by affinity chromatography, Epitheorese Klin. Farmakol. Farmakokinet. 5(2): 87-102 (1991) | |
| Publication Date | Received for publication: 25 July 1991
Accepted for publication: 30 August 1991 |
|
| Full Text Language | English | |
| Order – Buy | Ηλεκτρονική Μορφή: pdf (10 €) – Digital Type: pdf (10 €)
pharmakonpress[at]pharmakonpress[.]gr |
|
| Keywords | TSF, human pituitaries, affinity chromatography. | |
| Other Terms | review article | |
| Summary | A rather simple and efficient method has been developed for separation and purification of TSH from aceton dried human pituitary glands. The major portion of FSH, LH and TSH activities are obtained in a separate fraction after acetone and ethanol extraction and Sephadex gel filtration chromatography. TSH and FSH were separated from LH by adsorption on DEAE-Cellulose with 0.01 M glycine buffer at pH 9.5, and elution with gradient to 0.3 M. Highly specific anti-TSH antibodies were purified from a crude antiserum preparation by affinity chromatograpy using HCG-Sepharose column as the absorbant. This procedure eliminated completely the original LH binding activities. The antibody was then coupled to CNBr-Sepharose 4B and used in separating a highly purified TSH from FSH. TSH was labelled with, 125I (Chloramine T method) for identification. The eluates of affinity chromatography and iodination were assayed for TSH, FSH and LH; no FSH or LH activities were present in the TSH fractions. The purified TSH was then used for the immunization of rabbits. The antisera generated were of satisfactory specificity and titre. | |
| References | 1. Parlow, A.F., Condliffe, P.G., Reichert, L.E. Jr., Wilhelmi, A.E.: Recovery and partial purification of FSH and LH during the purification of TSH from human pituitary glands. Endocrinology 76: 27 (1965)
2. Parlow, A.F., Wilhetmi, A.E., Reichert, L.E.: Further studies on the fractionation of human pituitary glands. Endocrinology 77: 1126 (1965) 3. Wilhelmi, A.E.: Fractionation of human pituitary glands. Canad. J. Biochem. Physiol. 39: 1659 (1961) 4. Ackers, G.K.: Analytical gel chromatography of proteins. Adv. Protein. Chem. 24: 343 (1970) 5. Determann, H., Brewer, J.E.: Gel chromatography. In: Chromatography (E. Heftmann ed.), p. 362, 3rd edition. Reinhold, New York, 1975 6. Gel Filtration, Theory and Practice. Pharmacia Fine Chemicals. Uppsala, Sweden, p. 7, 1982-1983 7. Johnston, H.H.: Production of antisera to polypeptide hormones. In: Hormones in Human Blood. Detection and Assay (H.N. Antoniades, ed.), p. 65, Harvard University Press, Cambridge and London, England, 1976 8. Peterson, E.A., Sober, H.A.: Chromatography of proteins I. Cellulose ion-exchange adsorbents. J. Am. Chem. Soc. 78: 751 (1956) 9. Ion-exchange Chromatography. Principles and Methods. Pharmacie Fine Chemicals, Uppsala, Sweden, 1981 10. Axen, R., Porath, J., Ernback, S.: Chemical coupling of pleptides and proteins to polysaccharides by means of cyanogen halides. Nature 214: 1302 (1967) 11. Affinity Chromatography, Principles and Methods. Pharmacia Fine Chemicals. Uppsala, Sweden, 1979 12. Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.J.: Biol. Chem. 193: 265 (1951) 13. Hunter, W.M., Greenwood, F.C.: Preparation of iodine-131 labelled hGH of high specific activity. Nature 194: 495 (1962) 14. Hartree, A.S.: Separation and partial purification of protein hormones from pituitary glands. Biochem. J. 100: 754 (1966) 15. Torjesen, P.A., Sand, T., Noiman, N., Trygstad, O., Foss, I.: Isolation of LH, FSH and TSH from human pituitaries after the removal of hGH. Acta Endocrinologica 77: 485 (1974) 16. Sairam, M.R., Li, C.H.: Human pituitary thyrotrophin. Isolation and characterization of the hormone and its subunits. Can. J. Biochem. 55: 747 (1977) 17. Bates, R.W., Garrison, M.M., Cooper, J.A., Condliffe, P.G.: Further studies on the purification of human thyrotropin. Endocrinology 83: 721 (1968) 18. Condliffe, P.G.: Purification of human hyrotrophin. Endocrinology 72: 893 (1963) 19. Chapman, E.C., Renwick, G.C., Livesey, J.H.: The isolation of human pituitary hormones from frozen glands. J. Clin. Endocrinol. Metab. 53: 1008 (1981) 20. Lock, A.J., Janneke van Denderen, Aarden, L.A.: Purification for bovine thyroid-stimulating hormone by a monoclonal antibody. Endocrinology 122: 319 (1988) |
|
| Relative Papers |
Online ISSN 1011-6575
Άρθρα Δημοσιευμένα σε αυτό το Περιοδικό Καταχωρούνται στα:
- Chemical Abstracts
- Elsevier’s Bibliographic Databases: Scopus, EMBASE, EMBiology, Elsevier BIOBASE SCImago Journal and Country Rank Factor
Articles published in this Journal are Indexed or Abstracted in: • Chemical Abstracts • Elsevier’s Bibliographic Databases: Scopus, EMBASE, EMBiology, Elsevier BIOBASE SCImago Journal and Country Rank Factor
Συντακτικη Επιτροπή-Editorial Board
| ΕΤΗΣΙΑ ΣΥΝΔΡΟΜΗ 1991 – ANNUAL SUBSCRIPTION 1991 | |
| Γλώσσα Πλήρους Κειμένου – Full Text Language | Αγγλικά, Γαλλικά – English, French |
| Παραγγελία – Αγορά – Order – Buy | Ηλεκτρονική Μορφή: pdf (70 €) – Digital Type: pdf (70 €)
pharmakonpress[at]pharmakonpress[.]gr |
| Έντυπη Μορφή (70 € + έξοδα αποστολής) – Printed Type (70 € + shipping)
pharmakonpress[at]pharmakonpress[.]gr |
|