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In-vitro Mutagenesis

In-vitro Mutagenesis

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Related Experiment Video

Updated: Feb 28, 2026

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

Computationally optimized deimmunization libraries yield highly mutated enzymes with low immunogenicity and enhanced

Regina S Salvat¹, Deeptak Verma², Andrew S Parker²

¹Thayer School of Engineering, Dartmouth College, Hanover, NH 03755.

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

|June 14, 2017

Summary

We developed an algorithm to reduce immune responses to therapeutic proteins by optimizing mutations. This approach successfully created deimmunized protein variants with high efficacy and reduced immunogenicity, improving potential treatments.

Keywords:

T-cell epitope combinatorial library computational protein design deimmunization immunogenicity

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Last Updated: Feb 28, 2026

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

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Assessing Somatic Hypermutation in Ramos B Cells after Overexpression or Knockdown of Specific Genes

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Published on: November 1, 2011

Area of Science:

Biotechnology
Immunology
Protein Engineering

Background:

Therapeutic proteins are vital for disease treatment but can trigger harmful immune responses.
Immune surveillance of biotherapeutics can lead to reduced efficacy and safety concerns.

Purpose of the Study:

To develop a method for eliminating T-cell epitopes in biotherapeutics.
To mitigate detrimental immune recognition of therapeutic proteins.
To create deimmunized protein variants with retained or improved function.

Main Methods:

Developed a Pareto optimal deimmunization library design algorithm.
Optimized protein libraries considering simultaneous effects of mutations on function and epitope content.
Utilized high-throughput screening and ex vivo cellular immunoassays.

Main Results:

Screening of a 10-site library of P99 β-lactamase (P99βL) showed high fitness and reduced immunogenic potential.
A 30-site library yielded variants with enhanced activity and stability despite numerous deimmunizing mutations.
A 14-mutation variant silenced T-cell activation in 7 of 8 donors compared to wild-type P99βL.

Conclusions:

The multiobjective library design efficiently identified compatible epitope-deleting mutations.
The process generated highly active, aggressively deimmunized protein constructs.
This strategy offers a promising approach to enhance the safety and efficacy of protein therapeutics.