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US 4,228,068 AGranted
Peptide complexes of DNA-containing organisms
AssigneeR & Z Vermogensverwaltungsgesellschaft mbH
InventorGunter R. Wilhelm
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Abstract
DNA peptide complexes are prepared from organisms containing DNA to produce specific peptide complexes in a substantially pure state and useful for diagnostic, therapeutic and prophylactic purposes. The complexes can be prepared from bacteria, fungi, plants, human or animal tissue by homogination, centrification, absorption on DEAE cellulose, serial elution of the cellulose followed by dialysis and collection.
Claims3
The peptide complex, obtained from desoxyribonucleic acid-containing organisms, wherein
- (a) said organisms and parts or constituents thereof are homogenized in a native or denatured state in 0.2 M phosphate buffer (pH 7.2);
- (b) said homogenate is centrifuged;
- (c) the supernatant is stirred with phosphate buffer-loaded DEAE cellulose and charged into a column;
- (d) the loaded DEAE cellulose is eluted with 0.2 molar phosphate buffer until the absorption of the eluate at 280 nm is below 0.1, and thereafter further eluted with 0.1 M acetic acid-acetate solution (pH 3.2) until the absorption of the eluate at 280 nm is again below 0.1, thereupon eluted with a 3% NaCl containing 0.1 M acetic acid (pH 3.2) and the ribonucleic acid fraction (RNP) appearing in the eluate with the NaCl front is collected, dialyzed against water, concentrated and lyophilized, characterized in that
- I. the lyophilized RNP dissolved in water is mixed with phenol, heated to about 95.degree. to 100.degree. C. and, upon cooling, centrifuged until the phases are separated, the phenol phase is mixed with water and then repeatedly shaked out with ether, and the aqueous residue is lyophilized;
- or
- II. the RNP dissolved in water is subjected to high-voltage electrophoresis, and the peptide complex is isolated;
- or
- III. the peptide complex is isolated from the RNP dissolved in water, by thin film chromatography.
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