Showing papers on "Affinity chromatography published in 1969"
Journal Article•
TL;DR: The laboratory has been able to prepare highly efficient immunoadsorbents by coupling haptens or immunoglobulins to sepharose activated with cyanogen bromide (CNBr) using the general procedures reported by Cuatrecasas et al. (3) for selective enzyme purification by affinity chromatography.
Abstract: Our laboratory has been able to prepare highly efficient immunoadsorbents by coupling haptens or immunoglobulins to sepharose activated with cyanogen bromide (CNBr) (1–3). The general procedures reported by Cuatrecasas et al. (3) for selective enzyme purification by affinity chromatography are readily adapted to the specific purification of antibodies and antigens. These methods make the preparation of superior immunoadsorbents a remarkably easy task. Anti-hapten antibody purification . Sepharose activation is carried out as described (3); we have found it most satisfactory to add the sepharose slurry to a 25 mg/ml aqueous CNBr solution, immediately after adjusting the pH of the latter to 11.5, and the pH is maintained at 11.0 to 11.5 for the 8 to 10 min reaction time. Haptens with a free amino function are coupled to washed activated sepharose in 0.1 N sodium bicarbonate buffer, pH 9.0, at 4° to 5°C, also as previously described (3); we react 4 µmols of hapten/ml wet activated sepharose.
163 citations
TL;DR: Thyroxine-binding globulin has been separated from human serum by affinity chromatography using Sepharose to which l -thyroxine had been covalently attached and produced a single stained band on analytic disc-gel electrophoresis.
102 citations
TL;DR: 5′-(4-Aminophenyl-phosphoryl)-uridine-2′(3′)-phosphate was synthesized and coupled to Sepharose by activation with cyanogen bromide and used as a specific adsorbent for purification of ribonuclease A by affinity chromatography.
Abstract: 5′-(4-Aminophenyl-phosphoryl)-uridine-2′(3′)-phosphate was synthesized and coupled to Sepharose by activation with cyanogen bromide. This conjugate was used as a specific adsorbent for purification of ribonuclease A by affinity chromatography. Totally reduced and oxidized RNase preparations which are enzymatically inactive were not adsorbed or retarded by the RNase-specific column, while S-protein is strongly retarded.
96 citations
TL;DR: A two-fold increase in specific activity was achieved by this purification technique, and all of the non-proteolytic components were separated from the proteolytically active proteins.
66 citations
TL;DR: The "functional purification" of the crude, synthetic polypeptide by affinity chromatography was found to yield a synthetic fraction of greatly enhanced specific activity.
Abstract: The polypeptide corresponding to the amino acid sequence from residue 6 through 47 in staphylococcal nuclease has been synthesized by the solid-phase method. The synthetic product closely resembles the corresponding native polypeptide in both physical and chemical properties. The synthetic peptide may be recombined with the complimentary native peptide comprising residues 49 through 149 to form an active, semisynthetic enzyme. The “functional purification” of the crude, synthetic polypeptide by affinity chromatography was found to yield a synthetic fraction of greatly enhanced specific activity. This purification was accomplished on a column of Sepharose to which the complimentary native peptide had been covalently bound.
39 citations
TL;DR: An androstane derivative covalently bound to agarose was effective in removing testosterone binding globulin from plasma and the protein could subsequently be separated from the reagent in low yield by means of guanidine.
29 citations
TL;DR: The tyrosine-sensitive 3-deoxy-D- arabino -heptulosonate-7-phosphate (DAHP) synthetase was retarded by the column relative to the bulk of the protein and was purified about 100-fold, while the phenylalanine- sensitive enzyme was not retarded.
27 citations