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Related Experiment Video

Updated: Jun 21, 2026

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

Published on: January 17, 2015

Semisynthetic Phage Display Library Construction: Generation of Filtered Libraries.

Juan C Almagro¹, Mary Ann Pohl²

¹GlobalBio, Inc., Cambridge, Massachusetts 02138, USA juan.c.almagro@globalbioinc.com.

Cold Spring Harbor Protocols

|October 16, 2024

Summary

This study details a method to create therapeutic antibody discovery libraries. It focuses on selecting stable antibody fragments using M13 phage display and heat treatment for improved antibody discovery.

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Last Updated: Jun 21, 2026

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

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Published on: January 17, 2015

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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

Area of Science:

Biotechnology
Molecular Biology
Immunology

Background:

Antibody fragment display on M13 bacteriophages is key for identifying target-binding antibodies.
Therapeutic antibody discovery relies on robust and efficient library construction methods.

Purpose of the Study:

To describe the second step in constructing Antibody Libraries for Therapeutic Antibody Discovery (ALTHEA) Gold Plus Libraries.
To enhance antibody libraries by selecting for well-folded and thermostable single-chain variable fragments (scFvs).

Main Methods:

Displaying primary libraries (PLs) as scFv fusions to M13 phage protein pIII.
Implementing heat shock treatment and Protein L binding for selection of stable scFvs.
Removing unstable scFvs through washing and centrifugation, followed by quality assessment via clone sequencing.

Main Results:

Successfully constructed filtered libraries (FLs) enriched with thermostable scFvs.
Demonstrated the effectiveness of the filtration process in selecting high-quality antibody fragments.
Prepared libraries for the subsequent secondary library construction step.

Conclusions:

The described method effectively enriches antibody libraries with thermostable antibody fragments.
This filtration step is crucial for generating high-quality libraries for therapeutic antibody discovery.
The ALTHEA Gold Plus Libraries provide a foundation for advanced therapeutic antibody development.