Affinity Chromatography of Human Placental Alkaline Phosphatase

概要

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Human placental alkaline phosphatase (Alp) was purified by using the immunoadsorbent affinity chromatography. The commonly used procedures of purifying this enzyme were not only complicated but also time consuming as compared with this method, since the eluate of purified Alp could be easily obtained by only one step without further procedures. The anti-placental Alp antibody was prepared from a rabbit and coupled with CNBr-Sepharose 4B. The coupled complex was packed in a column. A crude extract of placental Alp was added and washed with 0.2M Tris-HCl buffer (pH 8.0) to exclude contaminated substances. The purified Alp was eluted from the column by 0.2M Na2CO3 solution (pH 11.4). A rechromatography of this column was also possible. The purity of the antigen (=purified Alp made by the previously reported method3) and the specificity of the antigen-antibody reaction are important for the purity of the eluate. To get a large volume of antibody from rabbits is difficult, which causes a limiting scale examination. Those points are to be improved.
Japan Society of Clinical Chemistryの論文