Abstract:
A novel biospecific affinity chromatographic procedure was developed for the purification of the sex steroid binding protein from Bufo arenarum. A charcoal column connected in series to the affinity column allows the removal of any ligand non-covalently bound to the matrix or released during its storage, thus avoiding the need for exhaustive and prolonged washing procedures. In addition, it is not necessary to remove the endogenous ligand from the starting material and the binding to the affinity column can be monitored to determine the time required to achieve the maximum yield. The advantages are the charcoal adsorption of the ligand "washed" from the affinity column by the protein to be purified and the amplification provided by the cyclic use of the system. The procedure improves the yield from less than 1% (by conventional procedures) to more than 50%. With minor modifications this procedure can be useful for the purification of binding proteins and receptors. © 1987.
Registro:
Documento: |
Artículo
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Título: | Cyclic biospecific affinity chromatographic method for the purification of the sex steroid binding protein (SBP): Application to the purification of SBP from toad |
Autor: | Santa-Coloma, T.A.; Garraffo, H.M.; Gros, E.G.; Charreau, E.H. |
Filiación: | Instituto de Biologia y Medicina Experimental, Obligado 2490, Buenos Aires, 1428, Argentina Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pab. 2, Buenos Aires, 1428, Argentina Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pab. 2, Buenos Aires, 1428, Argentina Instituto de Biologia y Medicina Experimental, Obligado 2490, Buenos Aires, 1428, Argentina Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pab. 2, Buenos Aires, 1428, Argentina
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Palabras clave: | sex hormone binding protein; affinity chromatography; animal cell; methodology; nonhuman; priority journal; protein purification; toad; Ammonium Sulfate; Androstane-3,17-diol; Animal; Bufo arenarum; Carrier Proteins; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Indicators and Reagents; Protein Binding; Sex Hormone-Binding Globulin; Stanolone |
Año: | 1987
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Volumen: | 415
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Número: | C
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Página de inicio: | 297
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Página de fin: | 304
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DOI: |
http://dx.doi.org/10.1016/S0378-4347(00)83221-1 |
Título revista: | Journal of Chromatography B: Biomedical Sciences and Applications
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Título revista abreviado: | J. Chromatogr. B Biomed. Sci. Appl.
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ISSN: | 03784347
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CODEN: | JCBAD
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CAS: | Ammonium Sulfate, 7783-20-2; Androstane-3,17-diol, 25126-76-5; Carrier Proteins; Indicators and Reagents; Sex Hormone-Binding Globulin; Stanolone, 521-18-6
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03784347_v415_nC_p297_SantaColoma |
Referencias:
- Mercier-Bodard, Alfsen, Baulieu, (1966) Proceedings of the Second Symposium on Steroid Hormones, Ghent, June 1965, International Congress Series 101, p. 212. , E. Exley, Excerpta Medica Foundation, Amsterdam
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- Iqbal, Johnson, Purification and characterization of human sex hormone binding globulin (1978) Journal of Steroid Biochemistry, 10, p. 535
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- Davis, (1964) Ann. N.Y. Acad. Sci., 121, p. 404
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- Sammons, Adams, Nishizawa, Ultrasensitive silver-based color staining of polypeptides in polyacrylamide gels (1981) Electrophoresis, 2, p. 135
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Citas:
---------- APA ----------
Santa-Coloma, T.A., Garraffo, H.M., Gros, E.G. & Charreau, E.H.
(1987)
. Cyclic biospecific affinity chromatographic method for the purification of the sex steroid binding protein (SBP): Application to the purification of SBP from toad. Journal of Chromatography B: Biomedical Sciences and Applications, 415(C), 297-304.
http://dx.doi.org/10.1016/S0378-4347(00)83221-1---------- CHICAGO ----------
Santa-Coloma, T.A., Garraffo, H.M., Gros, E.G., Charreau, E.H.
"Cyclic biospecific affinity chromatographic method for the purification of the sex steroid binding protein (SBP): Application to the purification of SBP from toad"
. Journal of Chromatography B: Biomedical Sciences and Applications 415, no. C
(1987) : 297-304.
http://dx.doi.org/10.1016/S0378-4347(00)83221-1---------- MLA ----------
Santa-Coloma, T.A., Garraffo, H.M., Gros, E.G., Charreau, E.H.
"Cyclic biospecific affinity chromatographic method for the purification of the sex steroid binding protein (SBP): Application to the purification of SBP from toad"
. Journal of Chromatography B: Biomedical Sciences and Applications, vol. 415, no. C, 1987, pp. 297-304.
http://dx.doi.org/10.1016/S0378-4347(00)83221-1---------- VANCOUVER ----------
Santa-Coloma, T.A., Garraffo, H.M., Gros, E.G., Charreau, E.H. Cyclic biospecific affinity chromatographic method for the purification of the sex steroid binding protein (SBP): Application to the purification of SBP from toad. J. Chromatogr. B Biomed. Sci. Appl. 1987;415(C):297-304.
http://dx.doi.org/10.1016/S0378-4347(00)83221-1