The affinity measurements were performed as described before [23 (link)]. In brief, ACE2-hFc was labeled by the Protein Labeling Kit RED-NHS 2nd Generation (NanoTemper) according to the manufacturer’s protocol. A degree of labeling (DOL) of < 3 was achieved and 10 nM of the labeled ACE2-hFc was applied in the measurements. Titration of the RBD variants was done by a Precision XS microplate sample processor (BioTek) in 384-well plates. Measurement was performed in Monolith (Nanotemper) using Monolith NT. Automated Capillary Chips (NanoTemper). The Excitation-Power was set to 40% and MST-Power to medium. The timeframe of 0.5 s up to 1.5 s was chosen to analyze the data by the MO Affinity Analysis software (NanoTemper) by Hill fit. For all RBD variants, a signal response above 18 and a signal to noise above 40 was obtained.
Schubert M., Bertoglio F., Steinke S., Heine P.A., Ynga-Durand M.A., Maass H., Sammartino J.C., Cassaniti I., Zuo F., Du L., Korn J., Milošević M., Wenzel E.V., Krstanović F., Polten S., Pribanić-Matešić M., Brizić I., Baldanti F., Hammarström L., Dübel S., Šustić A., Marcotte H., Strengert M., Protić A., Piralla A., Pan-Hammarström Q., Čičin-Šain L, & Hust M. (2022). Human serum from SARS-CoV-2-vaccinated and COVID-19 patients shows reduced binding to the RBD of SARS-CoV-2 Omicron variant. BMC Medicine, 20, 102.
Other organizations :
Technische Universität Braunschweig, Helmholtz Centre for Infection Research, Policlinico San Matteo Fondazione, Istituti di Ricovero e Cura a Carattere Scientifico, Karolinska Institutet, University of Rijeka, University of Pavia, Centre for Individualised Infection Medicine
Affinity measurements of ACE2-hFc binding to RBD variants
control variables
Degree of labeling (DOL) of ACE2-hFc was < 3
Concentration of labeled ACE2-hFc was 10 nM
Excitation-Power was set to 40%
MST-Power was set to medium
Timeframe of analysis was 0.5 s up to 1.5 s
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
