Calcium hydroxyapatite fractionation as a second ion exchange step in protein purification: Improvement of flow rate and resolution

The use of washed brushite, CaHPO 4, as a filter aid permitted the flow rates of calcium hydroxyapatite (HAP) columns to be made reliable and high. Regeneration of the column contents yielded pure, crystalline HAP, which could be mixed with fresh brushite and reused. Column fractionation over HAP-br...

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Published inAnalytical biochemistry Vol. 59; no. 1; pp. 16 - 23
Main Authors Skarstedt, Mark T., Bascomb, Shoshanna
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.05.1974
Subjects
Alcohol Oxidoreductases - isolation & purification
Aminohydrolases - isolation & purification
Asparaginase - isolation & purification
Calcium
Chromatography, DEAE-Cellulose
Chromatography, Ion Exchange
Cytidine
Escherichia - enzymology
Evaluation Studies as Topic
Hydroxyapatites
Klebsiella - enzymology
Methods
Proteins - isolation & purification
Saccharomyces cerevisiae - enzymology
Spectrophotometry, Ultraviolet
Time Factors
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Summary:The use of washed brushite, CaHPO 4, as a filter aid permitted the flow rates of calcium hydroxyapatite (HAP) columns to be made reliable and high. Regeneration of the column contents yielded pure, crystalline HAP, which could be mixed with fresh brushite and reused. Column fractionation over HAP-brushite with a phosphate gradient was used as a second ion exchange step in protein purification, the first step being fractionation over DEAE-cellulose. In two examples discussed, HAP-brushite fractionation in the presence of an effector molecule, a molecule for which the enzyme of interest had a specific affinity, gave marked improvement of resolution.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(74)90004-9