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Related Experiment Video

Updated: Feb 9, 2026

Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins

Enzymatic Modification and Flow Cytometry Assessment of Yeast Surface Displayed Proteins

Published on: May 30, 2025

Protein Engineering and Selection Using Yeast Surface Display.

Alessandro Angelini¹, Tiffany F Chen, Seymour de Picciotto

¹Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Building 76-261, 500 Main Street, Cambridge, MA, 02139, USA.

Methods in Molecular Biology (Clifton, N.J.)

|June 11, 2015

Summary

This summary is machine-generated.

This study details a yeast surface display method to isolate and engineer protein binders. The process uses magnetic beads and flow cytometry to enhance binder stability, affinity, and specificity for target proteins.

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Area of Science:

Biotechnology
Molecular Biology
Protein Engineering

Background:

Yeast surface display is a versatile platform for protein engineering.
Developing novel protein binders with specific biophysical properties is crucial for various applications.

Purpose of the Study:

To describe a protocol for de novo isolation of protein binders using yeast surface display.
To outline a method for enhancing the properties of isolated protein binders.

Main Methods:

Utilizing yeast surface display to present large combinatorial libraries.
Employing magnetic bead separation and fluorescence-activated cell sorting (flow cytometry) for binder selection.
Characterizing biophysical properties of selected clones.
Iterative rounds of mutagenesis and flow cytometry for property enhancement.

Main Results:

Successful isolation of protein binders from yeast surface display libraries.
Demonstrated enhancement of protein binder stability, affinity, and specificity.
Established a robust protocol for directed evolution of protein binders.

Conclusions:

Yeast surface display coupled with magnetic bead and flow cytometry selection is effective for isolating and engineering high-affinity protein binders.
This protocol provides a framework for developing tailored protein binders for specific research and therapeutic applications.