Estradiol binding synthetic polypeptides

Chemical Communications

Volumn , Issue 13, 2000, Pages 1135-1136

  Giraudi, G ^a   Giovannoli, C ^a   Tozzi, C ^a   Baggiani, C ^a   Anfossi, L ^a  

^a UNIVERSITY OF TURIN   (Italy)

Author keywords

[No Author keywords available]

Indexed keywords

ESTRADIOL; POLYPEPTIDE;

AMINO ACID COMPOSITION; ARTICLE; BINDING AFFINITY; HORMONE BINDING; IMMUNOPRECIPITATION; PEPTIDE SYNTHESIS; POLYMERIZATION; PROTEIN STRUCTURE;

EID: 0034617269     PISSN: 13597345     EISSN: None     Source Type: Journal    
DOI: 10.1039/a909798h     Document Type: Article

References (14)

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6. The polymerisation was performed in mixed aqueous/organic media (carbonate buffer 0.2 M pH 9/DMF, 1 + 1) using carbodiimide as a condensing agent. The mixtures were shaken according to the different reaction times at room temperature in the dark. Then the oligopeptide mixtures were filtered on 0.45 μm sintered glass.
7. We washed the resin with carbonate buffer (20 mM, pH 8.5) until the absorbance had returned to the baseline. After reversing the direction of the flow, the elution was performed by running a phosphate buffer (50 mM phosphate, 1 M NaCl, 1 mM EDTA, pH 7).
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10. 3 to check the signal of the labelled estradiol.
11. We added the oligopeptides at increasing concentrations to a constant amount of labelled estradiol, and after centrifugation in an Amicon MPS-1 device, equipped with YM1 membranes (nominal molecular weight cut-off of 1000 Da), we measured the ultrafiltered estradiol in the filtrate. The fraction of estradiol in the retentate plotted vs. the oligopeptide concentration allowed us to estimate the value of binding constants.
12. -3) to the amino acid solution, and then we performed the polymerisation as previously described. These quantities were proportional to the percentage of the corresponding amino acid in the polymerisation mixture and assured a significant measure of radioactivity. The small volumes added can be considered negligible in comparison to the volume of the whole polymerisation mixture.
13. We used antibodies directly against testosterone and tritium labelled testosterone for the affinity measurement of this steroid, and anti-progesterone antibodies and tritium labelled progesterone for the determination of the affinity for this one.
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