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Related Experiment Video

Updated: Feb 10, 2026

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Published on: May 1, 2018

Xenoprotein engineering via synthetic libraries.

Zachary P Gates¹, Alexander A Vinogradov², Anthony J Quartararo²

¹Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139 zgates@mit.edu blp@mit.edu.

Proceedings of the National Academy of Sciences of the United States of America

|May 23, 2018

Summary

This summary is machine-generated.

This study introduces a novel platform for engineering synthetic proteins with noncanonical amino acids. It enables rapid screening of protein libraries, leading to the de novo discovery of novel protein binders.

Keywords:

D-protein flow cytometry mirror-image miniprotein protein engineering xenoprotein

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Area of Science:

Chemical Biology
Protein Engineering
Synthetic Biology

Background:

Chemical synthesis enables creation of large, complex proteins.
Engineering synthetic proteins with noncanonical amino acids lags behind.
This capability is crucial for advancing protein science.

Purpose of the Study:

To develop a high-throughput platform for screening synthetic protein libraries.
To enable the engineering of synthetic proteins containing noncanonical amino acids.
To discover novel protein binders using a de novo approach.

Main Methods:

Utilized a one-bead one-compound protein library screening platform.
Employed magnetic bead enrichment and flow cytometry for efficient screening.
Applied high-throughput MS/MS sequencing for active compound identification.

Main Results:

Achieved screening throughput comparable to cell surface display methods.
Successfully performed de novo discovery of miniprotein binders.
Identified novel binders for a large protein target (approximately 150 kDa).

Conclusions:

The developed platform significantly advances synthetic protein engineering capabilities.
This method facilitates the discovery of novel protein binders, overcoming previous limitations.
Opens new avenues for designing functional synthetic proteins.