Purification of alkaline phosphatase by affinity chromatography, 1976

Robinson, Maxine Isaac

1970-1979

Affinity chromatography has been successfully employed for the purification of alkaline phosphatase (APase). A potential inhibitor of APase, 3-aminopropyl-phosphonic acid (3-APPA), was.coup1ed to agarose, Affi-Gel 10, to form an affinity adsorbent for APase. Phosphocellulose was used as a second adsorbent. A partially purified extract from C57BL mice was applied to both adsorbents in 0.1 M acetate buffer, pH 5.0, and eluted with 1.0 M KCl in 0.05 M tris buffer, pH 7.4. Both adsorbents were effective in adsorbing the enzyme, and the specific activity of the enzyme recovered from both adsorbents was greatly enhanced. The relative yields of the adsorbents differed greatly. Approximately 57% of the activity which was bound to the 3-aminopropylphosphonic acid-Affi-Gel 10 adsorbent was recovered in a 140-fold enrichment relative to the extract which was applied to the column. Only 13% of the bound activity could be eluted from the phosphocellulose adsorbent in a 125-fold enrichment. Because of the poor yield, the phosphocellulose adsorbent has been evaluated as noneffective for affinity chromatography purification of APase. Conversely, the 3-aminopropylphosphonic acid-Affi-Gel 10 adsorbent provides a rapid, one-step procedure for APase purification.

text

application/pdf

1976-07-01

thesis

Master of Science (MS)

Atlanta University

Biology

Lumb, Judith Rae

Clark Atlanta University

Georgia--Atlanta

http://hdl.handle.net/20.500.12322/cau.td:1976_robinson_maxine_i