Nucleic acids affinity purification

Affinity chromatography is similar in principle to the use of immuno-adsorbents for antibody and antigen purification (see Section IX, p. 375) and of insolubilized nucleic acids for purification of nucleic acids and related enzymes (see Section X, p. 384). The subject of affinity chromatography in general has been reviewed, ° and a mechanism... 

Macromolecules such as proteins, polysaccharides, nucleic acids differ only in their physicochemical properties within the individual groups and their isolation on the basis of these differences is therefore difficult and time consuming. Considerable decreases may occur during their isolation procedure due to denaturation, cleavage, enz3rmatic hydrolysis, etc. The ability to bind other molecules reversibly is one of the most important properties of these molecules. The formation of specific and reversible complexes of biological macromolecules can serve as basis of their separation, purification and analysis by the affinity chromatography [6]. 

Affinity chromatography can be applied to the isolation and purification of virtually all biological macromolecules. It has been used to purify nucleic acids, enzymes, transport proteins, antibodies, hormone receptor proteins, drug-binding proteins, neurotransmitter proteins, and many others. 

Gabrielsen, O. S., Homes, E., Komes, L., Ruet, A., and Oyen, T. B. (1989) Magnetic DNA affinity purification of yeast transcription factor T—a new purification principle for the ultrarapid isolation of near homogenous factor. Nucleic Acids Res. 17,6253-6267. 

Purification of exopolysaccharides is generally a difficult matter, owing to the high viscosity of most polymers. Quaternary ammonium compounds, which precipitate acidic polysaccharides, have been used successfully247 to separate acidic from neutral polysaccharides. Pretreatment with enzymes that preferentially cleave such undesirable contaminants as protein, nucleic acid, and cells results in isolation of improved products. Selection either of polysaccharide or of contaminating material on either ion-exchange or affinity-chromatog-... 

Affinity chromatography Has a wide number of uses and can be applied to the isolation and purification of virtually all biomolecules. Specific applications include nucleic acid purification, protein purification from cell and tissues extracts, and antibody purification from blood serum. There are a number of matrices used for the construct, and some examples of these and their uses are as follows heparin columns to separate cholesterol lipoproteins, lectin columns to separate carbohydrate groups, and phenyl boronate columns to separate glycated haemoglobins. 

Classical LLPC using aqueous-aqueous polymer systems based on Albertsson s [9] PEG-dextran system has provided a versatile tool for the separation of proteins and nucleic acids, thus increasing the arsenal of biopolymer purification methods currently dominated by gel filtration, ion-exchange chromatography, and affinity chromatography RPC. The technique operates... 

Many of the techniques used in protein purification procedures have also been adapted for use with nucleic acids. For example, several types of chromatography (e.g., ion-exchange, gel filtration, and affinity) have been used in several stages of nucleic acid purification and in the isolation of individual nucleic acid sequences. Because of its speed, HPLC has replaced many slower chromatographic separation techniques when small samples are involved. 

Proteins non-covalently adsorbed or covalently linked on solid-phase matrices have many applications in the wide area of science and technology (1 2). Indeed, in the field of separations science and technology, affinity selection based on the biospecific interaction of biomolecules with proteinaceous ligands immobilized on a solid-phase matrix has been widely noted as the separation method with highest selectivity, and is commonly utilized for separation and purification of biopolymers including proteins and nucleic acids (3,4). ... 

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