Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Yeast surface display for screening combinatorial polypeptide libraries

E T Boder1, K D Wittrup

  • 1Department of Chemical Engineering, University of Illinois, Urbana 61801, USA.

Nature Biotechnology
|June 1, 1997
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Isolation and characterization of human antibodies targeting human aspartyl (asparaginyl) beta-hydroxylase.

Human antibodies·2008
Same author

Substantial energetic improvement with minimal structural perturbation in a high affinity mutant antibody.

Journal of molecular biology·2004
Same author

Protein engineering by cell-surface display.

Current opinion in biotechnology·2001
Same author

High affinity T cell receptors from yeast display libraries block T cell activation by superantigens.

Journal of molecular biology·2001
Same author

Yeast surface display for directed evolution of protein expression, affinity, and stability.

Methods in enzymology·2000
Same author

The single cell as a microplate well.

Nature biotechnology·2000
Same journal

Author Correction: Sustained nitric oxide production by engineered E. coli remodels the tumor microenvironment and potentiates immunotherapy.

Nature biotechnology·2026
Same journal

Quantum computing in transition.

Nature biotechnology·2026
Same journal

Author Correction: Improved RNA base editing with guide RNAs mimicking highly edited endogenous ADAR substrates.

Nature biotechnology·2026
Same journal

Unlocking the chemical potential of filamentous fungi using prime editing.

Nature biotechnology·2026
Same journal

A genome-scale CRISPRi perturbation atlas of human induced pluripotent stem cells.

Nature biotechnology·2026
Same journal

Prime editing for precise genome engineering and modulation of fungal metabolism.

Nature biotechnology·2026
See all related articles

Yeast cell wall display enables engineering of mammalian proteins, improving antibody fragment selection and characterization. This eukaryotic system overcomes bacterial expression biases for enhanced protein folding and activity.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Background:

  • The yeast cell wall is a suitable platform for displaying eukaryotic proteins, including antibodies and receptors.
  • Endoplasmic reticulum-specific post-translational modifications are crucial for the proper folding and function of many mammalian proteins.
  • Bacterial expression systems can introduce biases in combinatorial libraries, potentially limiting protein diversity and function.

Purpose of the Study:

  • To leverage yeast cell wall display for engineering mammalian cell-surface and secreted proteins.
  • To utilize the Saccharomyces cerevisiae Aga2p receptor system for selecting improved antibody fragments.
  • To assess the utility of a eukaryotic host system for mitigating expression biases in protein libraries.

Main Methods:

Related Experiment Videos

  • Engineering mammalian proteins via C-terminal fusion to the Aga2p receptor on the yeast cell wall.
  • Utilizing random mutagenesis to create a library of single-chain variable fragment (scFv) antibody fragments.
  • Employing quantitative flow cytometry for fine discrimination of protein-ligand binding kinetics.

Main Results:

  • Demonstrated successful display of mammalian proteins on the yeast cell wall, facilitating endoplasmic reticulum-specific processing.
  • Selected scFv antibody fragments with a threefold decreased antigen dissociation rate compared to wild-type.
  • Showcased the advantage of a eukaryotic host system in reducing expression biases inherent in bacterial libraries.

Conclusions:

  • Yeast cell wall display is an effective strategy for engineering and selecting functional mammalian proteins, particularly antibodies.
  • The Aga2p fusion system coupled with flow cytometry provides a powerful method for optimizing protein binding kinetics.
  • Eukaryotic display systems offer a valuable alternative to bacterial systems for constructing and screening diverse protein libraries.