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Production of soluble ACE2 (sol-ACE2-Fc)

MH Markus Hoffmann
PA Prerna Arora
RG Rüdiger Groß
AS Alina Seidel
BH Bojan F. Hörnich
AH Alexander S. Hahn
NK Nadine Krüger
LG Luise Graichen
HH Heike Hofmann-Winkler
AK Amy Kempf
MW Martin S. Winkler
SS Sebastian Schulz
HJ Hans-Martin Jäck
BJ Bernd Jahrsdörfer
HS Hubert Schrezenmeier
MM Martin Müller
AK Alexander Kleger
JM Jan Münch
SP Stefan Pöhlmann
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293T cells were grown in a T-75 flask and transfected with 20 μg of sol-ACE2-Fc expression plasmid. At 10 h posttransfection, the medium was replaced and cells were further incubated for 38 h before the culture supernatant was collected and centrifuged (2,000 x g, 10 min, 4°C). Next, the clarified supernatant was loaded onto Vivaspin protein concentrator columns with a molecular weight cut-off of 30 kDa (Sartorius) and centrifuged at 4,000 x g, 4°C until the sample was concentrated by a factor of 20. The concentrated sol-ACE2-Fc was aliquoted and stored at −80°C until further use.

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