Author: Svenja Weiss; Jeromine Klingler; Catarina Hioe; Fatima Amanat; Ian Baine; Erna Milunka Kojic; Jonathan Stoever; Sean Liu; Denise Jurczyszak; Maria Bermudez-Gonzalez; Viviana Simon; Florian Krammer; Susan Zolla-Pazner
Title: A High Through-put Assay for Circulating Antibodies Directed against the S Protein of Severe Acute Respiratory Syndrome Corona virus 2 Document date: 2020_4_17
ID: 9iwjtwyx_11
Snippet: Luminex binding Ab assay. The SARS-CoV-2 antigens included a soluble recombinant trimerized form of the S protein and a recombinant RBD protein produced in Expi293F mammalian cells as described (mSpike, mRBD). 7 The antigens were covalently coupled individually to carboxylated xMAP beads at 4.0 μg protein/million beads using a two-step carbodiimide reaction with the xMAP Ab Coupling (AbC) Kit according to the manufacturers' instructions (Luminex.....
Document: Luminex binding Ab assay. The SARS-CoV-2 antigens included a soluble recombinant trimerized form of the S protein and a recombinant RBD protein produced in Expi293F mammalian cells as described (mSpike, mRBD). 7 The antigens were covalently coupled individually to carboxylated xMAP beads at 4.0 μg protein/million beads using a two-step carbodiimide reaction with the xMAP Ab Coupling (AbC) Kit according to the manufacturers' instructions (Luminex, Austin, TX). The coupled beads are pelleted, resuspended in storage buffer at 5 x 10 6 /mL (PBS, 0.1% bovine serum albumin (BSA), 0.02% Tween-20, and 0.05% sodium azide, pH 7.4), and stored at -80 o C. Three to five million beads were prepared in a single 1.5 mL conical tube.
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