Rhiel, Laura ; Krah, Simon ; Günther, Ralf ; Becker, Stefan ; Kolmar, Harald ; Hock, Björn (2024)
REAL-Select: Full-Length Antibody Display and Library Screening by Surface Capture on Yeast Cells.
In: PLOS ONE, 2014, 9 (12)
doi: 10.26083/tuprints-00027624
Artikel, Zweitveröffentlichung, Verlagsversion
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Kurzbeschreibung (Abstract)
We describe a novel approach named REAL-Select for the non-covalent display of IgG-molecules on the surface of yeast cells for the purpose of antibody engineering and selection. It relies on the capture of secreted native full-length antibodies on the cell surface via binding to an externally immobilized ZZ domain, which tightly binds antibody Fc. It is beneficial for high-throughput screening of yeast-displayed IgG-libraries during antibody discovery and development. In a model experiment, antibody-displaying yeast cells were isolated from a 1∶1,000,000 mixture with control cells confirming the maintenance of genotype-phenotype linkage. Antibodies with improved binding characteristics were obtained by affinity maturation using REAL-Select, demonstrating the ability of this system to display antibodies in their native form and to detect subtle changes in affinity by flow cytometry. The biotinylation of the cell surface followed by functionalization with a streptavidin-ZZ fusion protein is an approach that is independent of the genetic background of the antibody-producing host and therefore can be expected to be compatible with other eukaryotic expression hosts such as P. pastoris or mammalian cells.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2024 |
| Autor(en): | Rhiel, Laura ; Krah, Simon ; Günther, Ralf ; Becker, Stefan ; Kolmar, Harald ; Hock, Björn |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | REAL-Select: Full-Length Antibody Display and Library Screening by Surface Capture on Yeast Cells |
| Sprache: | Englisch |
| Publikationsjahr: | 19 August 2024 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2014 |
| Ort der Erstveröffentlichung: | San Francisco, California, US |
| Verlag: | PLOS |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | PLOS ONE |
| Jahrgang/Volume einer Zeitschrift: | 9 |
| (Heft-)Nummer: | 12 |
| Kollation: | 19 Seiten |
| DOI: | 10.26083/tuprints-00027624 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/27624 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichungsservice |
| Kurzbeschreibung (Abstract): | We describe a novel approach named REAL-Select for the non-covalent display of IgG-molecules on the surface of yeast cells for the purpose of antibody engineering and selection. It relies on the capture of secreted native full-length antibodies on the cell surface via binding to an externally immobilized ZZ domain, which tightly binds antibody Fc. It is beneficial for high-throughput screening of yeast-displayed IgG-libraries during antibody discovery and development. In a model experiment, antibody-displaying yeast cells were isolated from a 1∶1,000,000 mixture with control cells confirming the maintenance of genotype-phenotype linkage. Antibodies with improved binding characteristics were obtained by affinity maturation using REAL-Select, demonstrating the ability of this system to display antibodies in their native form and to detect subtle changes in affinity by flow cytometry. The biotinylation of the cell surface followed by functionalization with a streptavidin-ZZ fusion protein is an approach that is independent of the genetic background of the antibody-producing host and therefore can be expected to be compatible with other eukaryotic expression hosts such as P. pastoris or mammalian cells. |
| Freie Schlagworte: | Yeast, Saccharomyces cerevisiae, Library screening, Flow cytometry, Cloning, Cell fusion, Cell staining, Cell binding |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-276245 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut |
| Hinterlegungsdatum: | 19 Aug 2024 09:58 |
| Letzte Änderung: | 20 Aug 2024 06:16 |
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| Suche nach Titel in: | TUfind oder in Google |
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