scholarly journals Purification of foetal steroid-binding protein from human serum by affinity chromatography on 5α-androstane-3 β,17 β-diol 3-hemisuccinate-aminohexyl-Sepharose 6B

1986 ◽  
Vol 240 (1) ◽  
pp. 75-79 ◽  
Author(s):  
M L Wilkinson ◽  
M J Iqbal ◽  
A Forbes ◽  
T P Corbishley ◽  
R Williams
Keyword(s):  
Ligand Binding ◽  
Affinity Chromatography ◽  
Human Serum ◽  
Binding Protein ◽  
Gel Filtration ◽  
Specific Antiserum ◽  
Alkaline Ph ◽  
Salt Precipitation ◽  
Steroid Binding ◽  
Steroid Binding Protein

In order to develop an immunoassay for foetal steroid-binding protein in human serum, which is impossible to assay quantitatively in normal samples by conventional ligand-binding techniques, the protein was purified by salt precipitation, affinity chromatography and gel filtration. Elution was by competing ligand or alkaline pH. The purified protein was further characterized and a highly specific antiserum was raised in rabbits.

An enzyme-linked immunosorbent assay for fetal steroid binding protein of human serum

Steroids ◽  
1985 ◽  
Vol 45 (1) ◽  
pp. 31-38 ◽  
Author(s):  
M. Jawed Iqbal ◽  
Alastair Forbes ◽  
Timothy P. Corbishley ◽  
Mark L. Wilkinson ◽  
Roger Williams
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Steroid Binding ◽  
Steroid Binding Protein

The isolation, purification and some properties of pheromaxein, the pheromonal steroid-binding protein, in porcine submaxillary glands and saliva

1988 ◽  
Vol 118 (1) ◽  
pp. 47-NP ◽  
Author(s):  
W. D. Booth ◽  
C. A. White
Keyword(s):  
Liquid Chromatography ◽  
Affinity Chromatography ◽  
Sodium Dodecyl Sulphate ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Submaxillary Gland ◽  
Fast Protein Liquid Chromatography ◽  
Steroid Binding ◽  
Steroid Binding Protein

ABSTRACT Pheromaxein, the 16-androstene steroid-binding protein with a relative molecular mass of 15 000 was isolated in sub-milligram quantities from the submaxillary gland and saliva of the Gottingen miniature boar, after a fourfold purification involving the following methods: ultrafiltration for submaxillary gland cytosols and ethanol precipitation for saliva, Concanavalin-A-Sepharose affinity chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis, 'Extractigel-D' affinity chromatography (to remove sodium dodecyl sulphate) and fast protein-liquid chromatography. Yields of purified pheromaxein obtained after fast protein-liquid chromatography represented 10–20% of total protein present in an ultrafiltrate of a submaxillary gland cytosol. Fast protein-liquid chromatography separated the α- and β-charge isomers of pheromaxein which were shown to have isoelectric points of 4·78 and 5·35 respectively on flat-bed isoelectric focusing. Some data are provided for the variable occurrence of the isomeric forms of pheromaxein in relation to different breeds of pig. Five 16-unsaturated steroids showed the highest binding to pheromaxein. Other steroids of the 5α- and 5β-androstane series also showed some binding to pheromaxein, i.e. 17β-hydroxy-5α-androstan-3-one (19·2%), with 5α-androstan-3-one, which has a similar urinous odour to 5α-androst-16-en-3-one, showing the greatest binding (42·6%) relative to 5α-androst-16-en-3-one (100%). J. Endocr. (1988) 118, 47–57

scholarly journals Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein

2011 ◽  
Vol 11 (1) ◽  
pp. 64 ◽  
Author(s):  
Tiina A Riihimäki ◽  
Soili Hiltunen ◽  
Martina Rangl ◽  
Henri R Nordlund ◽  
Juha AE Määttä ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Binding Site ◽  
Binding Protein ◽  
Ligand Binding Site ◽  
Steroid Binding ◽  
Steroid Binding Protein

Interactions of foetal steroid binding protein with other binding proteins in human serum

1987 ◽  
Vol 169 (2-3) ◽  
pp. 299-308
Author(s):  
Alastair Forbes ◽  
Mark L. Wilkinson ◽  
M.Jawed Iqbal ◽  
Roger Williams
Keyword(s):  
Human Serum ◽  
Binding Proteins ◽  
Binding Protein ◽  
Steroid Binding ◽  
Steroid Binding Protein

Purification of the sex steroid binding protein from human serum

Biochemistry ◽  
1975 ◽  
Vol 14 (5) ◽  
pp. 957-963 ◽  
Author(s):  
Kenneth E. Mickelson ◽  
Philip H. Petra
Keyword(s):  
Human Serum ◽  
Binding Protein ◽  
Sex Steroid ◽  
Steroid Binding ◽  
Steroid Binding Protein

The Different Forms of Antithrombin III in Serum

1977 ◽  
Vol 38 (02) ◽  
pp. 0494-0503 ◽  
Author(s):  
D. S Pepper ◽  
D Banhegyi ◽  
J. D Cash
Keyword(s):  
Molecular Weight ◽  
Affinity Chromatography ◽  
Human Serum ◽  
Degradation Product ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Free Form ◽  
Polymeric Form ◽  
At Iii ◽  
Human Thrombin

SummaryAntithrombin III (AT III) complexes were isolated from human serum by affinity chromatography and gel filtration. In the first step of the preparation, using heparin-agarose chromatography, we observed that the complexed form of AT III bound less strongly to the gel than the free form and that about half of the AT III was free. With further purification a 2.5 × 105 molecular weight complex was isolated. Using 125I labelled human thrombin, this complex was radioactive indicating the presence of thrombin. Only in a synthetic thrombin-AT III system was a 9 × 104 molecular weight complex detected, but not in serum. These facts suggest that in serum AT III complexes may exist in a polymeric form. Also, an AT III antigen derived from the original AT III molecule, but not complexed, was isolated which may be a degradation product.Abbreviations used: AT-III, antithrombin III. Hepes, N-2-Hydroxyethylpiperazine-N-2-Ethanesulphonic acid.

Foetal steroid binding protein in British and Japanese women

1987 ◽  
Vol 114 (4) ◽  
pp. 584-588 ◽  
Author(s):  
M. Jawed Iqbal ◽  
Alastair Forbes ◽  
Mark L. Wilkinson ◽  
John W. Moore ◽  
Roger Williams ◽  
...  
Keyword(s):  
Ovarian Function ◽  
Binding Protein ◽  
Premenopausal Women ◽  
Japanese Women ◽  
Sex Hormone Binding Globulin ◽  
Partial Purification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Binding Characteristics ◽  
Steroid Binding ◽  
Steroid Binding Protein

Abstract. In order to examine the newly-discovered sex-steroid binding protein, foetal steroid binding protein (FSBP) in different populations, its binding characteristics and its level were studied by two-tier column ligand binding assay and enzyme-linked immunosorbent assay (ELISA) respectively. In 10 Japanese premenopausal women, analysis of 5α-dihydrotestosterone (DHT) binding in the Cibacron Blue 3GA-Sepharose 6B portion of the column showed a rising plateau pattern with a mean maximum binding of 31.1 ± 7.41%, whereas of 9 similar British women, 8 displayed unsaturable, non-cooperative binding of 11.6 ± 8.22% (P < 0.01). After partial purification of FSBP in these samples, the protein exhibited saturable binding kinetics, median binding 25 (interquartiles 23–34) and 19 (13–25) nmol DHT/l in Japanese and British women, respectively (P < 0.05). By analyzing FSBP by ELISA in 56 Japanese (45 premenopausal) and 59 British (25 premenopausal) women, higher levels were obtained in the whole Japanese group (P = 0.0016) and in the premenopausal Japanese women (P = 0.018) than in their British counterparts. In both nationalities, FSBP levels were higher in premenopausal women, and there was a significant negative correlation of FSBP with age in both populations, particularly in postmenopausal women. FSBP levels did not correlate with weight, parity, sex hormone binding globulin or albumin levels. The influence of FSBP on free steroid levels remains unclear, but some relationship with ovarian function seems a possibility.

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