Krištof Bozovičar (Avtor), Barbara Jenko (Avtor), Anže Meden (Avtor), Jernej Kovač (Avtor), Tomaž Bratkovič (Avtor)

Povzetek

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.

Ključne besede

peptidne knjižnice;ELISA;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: UL FFA - Fakulteta za farmacijo
UDK: 543.544+577.27
COBISS: 65576707 Povezava se bo odprla v novem oknu
ISSN: 2045-2322
Št. ogledov: 20
Št. prenosov: 10
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarne ključne besede: Imunoglobulini;Kromatografija;Analizna kemija;
Vrsta dela (COBISS): Članek v reviji
Strani: str. 1-12
Letnik: ǂVol. ǂ11
Zvezek: ǂart. no. ǂ11650
Čas izdaje: 2021
DOI: 10.1038/s41598-021-91208-0
ID: 18434263