Author: Wysocki, Jan; Schulze, Arndt; Batlle, Daniel
Title: Novel Variants of Angiotensin Converting Enzyme-2 of Shorter Molecular Size to Target the Kidney Renin Angiotensin System Document date: 2019_12_17
ID: 0vozochc_9
Snippet: Two short and enzymatically active ACE2 variants were selected for large scale production and purification. From stably transfected HEK cells, single clones were selected and expanded to 220 mL flasks. Conditioned serum-free medium from a clone of stably transfected HEK 293 cells that overexpressed the two short ACE2 variants was subjected to anion exchange Q-column on AKTA chromatography system in 25 mM Tris-HCl, pH 8.0 and eluted by applying in.....
Document: Two short and enzymatically active ACE2 variants were selected for large scale production and purification. From stably transfected HEK cells, single clones were selected and expanded to 220 mL flasks. Conditioned serum-free medium from a clone of stably transfected HEK 293 cells that overexpressed the two short ACE2 variants was subjected to anion exchange Q-column on AKTA chromatography system in 25 mM Tris-HCl, pH 8.0 and eluted by applying increasing concentration of NaCl. Eluted fractions were screened for ACE2 activity was using Mca-APK-Dnp substrate. Fractions containing ACE2 activity were applied to SDS-PAGE, transferred to PVDF membrane and stained with Brilliant Blue to assess protein purity (Supplementary Materials Figure S2 , showing ACE2-1-619 as an example).
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