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  • Focused peptide library screening as a route to a superior affinity ligand for antibody purification [Elektronski vir]
    Bozovičar, Krištof ...
    Affinity chromatography is the linchpin of antibody downstream processing and typically relies on bacterial immunoglobulin (Ig)-binding proteins, epitomized by staphylococcal protein A-based ligands. ... However, such afnity ligands are fairly costly and sufer from chemical instability, leading to ligand denaturation and leaching from chromatographic support. Innovations in this area are aimed at developing robust and highly selective antibody ligands capable of withstanding harsh column sanitization conditions. We report the development and frst-stage characterization of a selective short linear peptide ligand of the IgG Fc region capable of capturing all four IgG subclasses. The ligand was discovered through in vitro directed evolution. A focused phage-display library based on a previously identifed peptide lead was subjected to a single-round screen against a pool of human IgG. The hits were identifed with next-generation sequencing and ranked according to the enrichment ratio relative to their frequency in the pre-screened library. The top enriched peptide GSYWYNVWF displaying highest afnity for IgG was coupled to bromohydrin-activated agarose beads via a branched linker. The resulting afnity matrix was characterized with a dynamic binding capacity of approx. 43 mg/mL, on par with commercially employed protein A-based resin.
    Source: Scientific reports [Elektronski vir]. - ISSN 2045-2322 (Vol. 11, art. no. 11650, 2021, str. 1-12)
    Type of material - e-article ; adult, serious
    Publish date - 2021
    Language - english
    COBISS.SI-ID - 65576707

    Link(s):

    https://www.nature.com/articles/s41598-021-91208-0
    Repository of the University of Ljubljana – RUL
    DOI

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