ArticleMethods in molecular biology (Clifton, N.J.)2023
Antibody Affinity and Stability Maturation by Error-Prone PCR.
Article in Methods in molecular biology (Clifton, N.J.), 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
What it found
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Who cites it
5 citing papers in PubMed.
- Generation of a CDR H3 Randomized Monoclonal Antibody Library by Kunkel Mutagenesis.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Deaminase-driven random mutation enables efficient DNA mutagenesis for protein evolution.Chemical science · 2025Article
- Development of an optimized cell-based selection system for phage display libraries.Biology methods & protocols · 2025Article
- Development and experimental validation of computational methods for human antibody affinity enhancement.Briefings in bioinformatics · 2024Article
- Construction of a Human Immune Library from Gallbladder Cancer Patients for the Single-Chain Fragment Variable (Antibodies (Basel, Switzerland) · 2024Article
Corrections and comments
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Authors and funding
7 authors.
Funding
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Abstract
Human antibodies are the most important class of biologicals, and antibodies - human and nonhuman - are indispensable as research agents and for diagnostic assays. When generating antibodies, they sometimes show the desired specificity profile but lack sufficient affinity for the desired application. In this article, a phage display-based method and protocol to increase the affinity of recombinant antibody fragments is given.The given protocol starts with the construction of a mutated antibody gene library by error-prone PCR. Subsequently, the selection of high-affinity variants is performed by panning on immobilized antigen with washing conditions optimized for off-rate-dependent selection. A screening ELISA protocol to identify antibodies with improved affinity and an additional protocol to select antibodies with improved thermal stability is described.
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Registered trials
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