Purification and Tryptic Digestion of SARS-CoV-2 Antibodies

Anti-S IgG was captured through affinity purification with recombinant trimerized S protein-coated Maxisorp NUNC-Immuno plates (Thermo Fisher Scientific, Roskilde, Denmark) (18 (link)), while total IgG was enriched using protein G Sepharose Fast Flow 4 beads (32 (link)). A 100 mM formic acid solution was used for antibody elution, followed by sample drying through vacuum centrifugation and reconstitution in 25 mM ammonium bicarbonate. The purified antibodies were subjected to tryptic digestion to obtain glycopeptides, as described previously (29 (link), 32 (link)). For the samples from Brazil, a minimum of 2 Visucon-F standards, 4 pooled anti-S IgG samples and 2 blanks were included per plate. For the German samples at least 1 Visucon-F standard and 1 blank was included on each plate.

Free full text: Click here

Siekman S.L., Pongracz T., Wang W., Nouta J., Kremsner P.G., da Silva-Neto P.V., Esen M., Kreidenweiss A., Held J., Trapé Á.A., Fendel R., de Miranda Santos I.K, & Wuhrer M. (2022). The IgG glycome of SARS-CoV-2 infected individuals reflects disease course and severity. Frontiers in Immunology, 13, 993354.

Publication 2022

Maxisorp NUNC-Immuno plates

Affinity purification Ammonium bicarbonate Anti igg Antibodies Antibody Centrifugation Digestion Formic acid Glycopeptides Protein g Recombinant protein Sepharose Tryptic Vacuum

Corresponding Organization : Leiden University Medical Center

Other organizations : University of Tübingen, Centre de Recherche Médicales de Lambaréné, German Center for Infection Research, Universidade de São Paulo

Top 5 similar protocols

Variable analysis

independent variables

Affinity purification with recombinant trimerized S protein-coated Maxisorp NUNC-Immuno plates to capture Anti-S IgG
Protein G Sepharose Fast Flow 4 beads to enrich total IgG
Tryptic digestion to obtain glycopeptides

dependent variables

Purified antibodies

control variables

Visucon-F standards
Pooled anti-S IgG samples
Blanks

positive controls

Visucon-F standards
Pooled anti-S IgG samples

negative controls

Blanks

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?

Revolutionizing how scientists
search and build protocols!