A new polymer supported packing material as a substitute of hydroxylapatite: Purification of Topoisomerases
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Hydroxylapatite (HA), a form of calcium phosphate, finds extensive usages in the fractionation and purification of proteins, enzymes and nucleic acids. However, commercial HA preparations for liquid chromatography (LC) differ from many other LC packing materials in that the former is not polymer supported. We have now developed a substitute packing material which uses Fractosil 200 as the polymer support. Other polymer supports can also be similarly used. Being Fractosil 200 supported, this Hydroxylapatite Substitute (HAS) matrix can be employed for open column LC as well as for HPLC purposes. Although the exact chemical nature of HAS is different from the conventional HA, HAS mimicks the functions of the latter support. Due to the high’ binding capacity of HA many DNA binding proteins have been partially purified on this support. In the present work HAS has been successfully used in the partial purification of Topoisomerase I and II from wheat germ.
Key WordsLiquid chromatography Hydroxylapatite substitute, HAS Protein purification Topoisomerases
List of Common Abbreviations
Bovine serum albumin
40 mM Tris-acetate, 2 mM EDTA
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