Directed Evolution of Binding Proteins by Cell Surface Display: Analysis of the Screening Process

2003 ◽  
pp. 111-126 ◽  
Author(s):  
K. Dane Wittrup
Keyword(s):  
Cell Surface ◽  
Directed Evolution ◽  
Binding Proteins ◽  
Surface Display ◽  
Cell Surface Display ◽  
Screening Process

scholarly journals Development of a yeast cell surface display method using the SpyTag/SpyCatcher system

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kaho Kajiwara ◽  
Wataru Aoki ◽  
Naoki Koike ◽  
Mitsuyoshi Ueda
Keyword(s):  
Amino Acids ◽  
Cell Surface ◽  
Yeast Cell ◽  
Directed Evolution ◽  
Surface Display ◽  
Cell Surface Display ◽  
Protein Ligation ◽  
Continuous Evolution ◽  
Yeast Cell Surface

AbstractYeast cell surface display (YSD) has been used to engineer various proteins, including antibodies. Directed evolution, which subjects a gene to iterative rounds of mutagenesis, selection and amplification, is useful for protein engineering. In vivo continuous mutagenesis, which continuously diversifies target genes in the host cell, is a promising tool for accelerating directed evolution. However, combining in vivo continuous evolution and YSD is difficult because mutations in the gene encoding the anchor proteins may inhibit the display of target proteins on the cell surface. In this study, we have developed a modified YSD method that utilises SpyTag/SpyCatcher-based in vivo protein ligation. A nanobody fused with a SpyTag of 16 amino acids and an anchor protein fused with a SpyCatcher of 113 amino acids are encoded by separate gene cassettes and then assembled via isopeptide bond formation. This system achieved a high display efficiency of more than 90%, no intercellular protein ligation events, and the enrichment of target cells by cell sorting. These results suggested that our system demonstrates comparable performance with conventional YSD methods; therefore, it can be an appropriate platform to be integrated with in vivo continuous evolution.

scholarly journals Secretion-and-capture cell-surface display for selection of target-binding proteins

2011 ◽  
Vol 24 (6) ◽  
pp. 525-530 ◽  
Author(s):  
J. A. Rakestraw ◽  
D. Aird ◽  
P. M. Aha ◽  
B. M. Baynes ◽  
D. Lipovsek
Keyword(s):  
Cell Surface ◽  
Binding Proteins ◽  
Surface Display ◽  
Cell Surface Display ◽  
Target Binding ◽  
Selection Of

scholarly journals Directed Evolution of an Angiopoietin-2 Ligand Trap by Somatic Hypermutation and Cell Surface Display

2013 ◽  
Vol 288 (46) ◽  
pp. 33205-33212 ◽  
Author(s):  
Nicholas P. J. Brindle ◽  
Julian E. Sale ◽  
Hiroshi Arakawa ◽  
Jean-Marie Buerstedde ◽  
Teonchit Nuamchit ◽  
...  
Keyword(s):  
Cell Surface ◽  
Directed Evolution ◽  
Somatic Hypermutation ◽  
Surface Display ◽  
Cell Surface Display ◽  
Angiopoietin 2

scholarly journals Directed Evolution of a Lysosomal Enzyme with Enhanced Activity at Neutral pH by Mammalian Cell-Surface Display

2008 ◽  
Vol 15 (12) ◽  
pp. 1277-1286 ◽  
Author(s):  
Kai-Chuan Chen ◽  
Chia-Hung Wu ◽  
Chuan-Yuan Chang ◽  
Wei-Cheng Lu ◽  
Qingzong Tseng ◽  
...  
Keyword(s):  
Cell Surface ◽  
Directed Evolution ◽  
Mammalian Cell ◽  
Lysosomal Enzyme ◽  
Surface Display ◽  
Cell Surface Display ◽  
Neutral Ph ◽  
Enhanced Activity

Ligand fishing based on cell surface display of enzymes for inhibitor screening

2021 ◽  
pp. 338359
Author(s):  
Yun-Cong Yuan ◽  
Xiao-Lin Bai ◽  
Yi-Ming Liu ◽  
Xiao-Yue Tang ◽  
Hao Yuan ◽  
...  
Keyword(s):  
Cell Surface ◽  
Surface Display ◽  
Cell Surface Display ◽  
Inhibitor Screening ◽  
Ligand Fishing

scholarly journals Cover Feature: Bacterial Cell‐Surface Display of Semisynthetic Cyclic Peptides (ChemBioChem 1/2019)

ChemBioChem ◽  
2018 ◽  
Vol 20 (1) ◽  
pp. 4-4
Author(s):  
Shubhendu Palei ◽  
Kira S. Becher ◽  
Christian Nienberg ◽  
Joachim Jose ◽  
Henning D. Mootz
Keyword(s):  
Cell Surface ◽  
Bacterial Cell ◽  
Cyclic Peptides ◽  
Surface Display ◽  
Cell Surface Display ◽  
Bacterial Cell Surface
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