REAL-Select: full-length antibody display and library screening by surface capture on yeast cells.
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 immobi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4264850?pdf=render |
| _version_ | 1818172484267016192 |
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| author | Laura Rhiel Simon Krah Ralf Günther Stefan Becker Harald Kolmar Björn Hock |
| author_facet | Laura Rhiel Simon Krah Ralf Günther Stefan Becker Harald Kolmar Björn Hock |
| author_sort | Laura Rhiel |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T19:13:21Z |
| format | Article |
| id | doaj.art-389150e3d90a4193821785b3994a8b80 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T19:13:21Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-389150e3d90a4193821785b3994a8b802022-12-22T00:53:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01912e11488710.1371/journal.pone.0114887REAL-Select: full-length antibody display and library screening by surface capture on yeast cells.Laura RhielSimon KrahRalf GüntherStefan BeckerHarald KolmarBjörn HockWe 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.http://europepmc.org/articles/PMC4264850?pdf=render |
| spellingShingle | Laura Rhiel Simon Krah Ralf Günther Stefan Becker Harald Kolmar Björn Hock REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. PLoS ONE |
| title | REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. |
| title_full | REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. |
| title_fullStr | REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. |
| title_full_unstemmed | REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. |
| title_short | REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. |
| title_sort | real select full length antibody display and library screening by surface capture on yeast cells |
| url | http://europepmc.org/articles/PMC4264850?pdf=render |
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