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Large-scale purification of angiotensin I-converting enzyme from human plasma utilizing an immunoadsorbent affinity gel

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
103
Issue
2
Identifiers
DOI: 10.1016/0003-2697(80)90630-2
Keywords
  • Purification Of Enzymes And Proteins
Disciplines
  • Biology

Abstract

Abstract Angiotensin I-converting enzyme was purified 1500-fold from human plasma utilizing an immunoadsorbent affinity gel prepared by coupling antibody to baboon lung angiotensin I-converting enzyme to CNBr-activated Sepharose 4B. The enzyme was eluted from the gel using 2 m magnesium chloride, pH 5.8. Subsequent hydroxylapatite and Sephadex G-200 chromatography yielded 2.6 mg of homogenous enzyme with a specific activity of 40 units/mg with hippuryl- l-histidyl- l-leucine as substrate from 48 liters of plasma. Use of the immunoadsorbent allowed the 48 liters of plasma to be processed in one-half the time it previously took to process 2 liters of plasma by other methods. This protocol enables us to obtain sufficient amounts of enzyme for structural studies that were previously impossible because of insufficient amounts of enzyme.

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