Affinity Chromatography - download pdf or read online

By Jaroslava Turková (Eds.)

ISBN-10: 0444416056

ISBN-13: 9780444416056

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8. 2 Cooperative adsorption column chromatography Fig. 9 shows typical elution profiles of oligoadenylic acid at various temperatures. The elution centre of the peak is shifted and the peak shape is distorted from a Gaussian type by a decrease in temperature from 15 to O°C. These results indicate that the binding of oligoadenylic acid to polyuridylic acid is stabilized by decreasing the temperature. Further, the distorted elution profiles indicate the cooperativity of the binding of oligoadenylic acid to polyuridylic acid.

M. Chaiken, Biochemistry, 14 (1975) 2343-2349. - 31 ELUTION ANALYSIS The validity of this equation was checked by Dunn and Chaiken (1975) by examining the chromatography of staphylococcal nuclease on thymidine 3’+-Sepharose-aminophenyl phosphate) 5’-phosphate (abbreviation: pdTpAP-Sepharose) as a function of the concentration of both the immobilized nucleotide and the soluble nucleotide used for the elution of nuclease. Fig. 1 shows the effect of the dilution of the affinity matrix with unsubstituted Sepharose.

The scatter in KL seems to be independent of the nature of the substituents bound to thymidine. The agreement of the KL values (within experimental error) shows that the nuclease-affinity matrix interaction is independent of the presence of another ligand. In contrast to the KL values, the values of KI are strongly dependent on the substitution of the ligand. Virtually identical KI values for pdTp and pdTpAP indicate that the aminophenyl group of the spacer arm has no effect on the interaction of nuclease with the immobilized nucleotide.

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Affinity Chromatography by Jaroslava Turková (Eds.)

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