ArticleProceedings of the National Academy of Sciences of the United States of America2000
Quantitative analysis of the effect of the mutation frequency on the affinity maturation of single chain Fv antibodies.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 71 papers, 1 of them a synthesis that pooled it.
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Who cites it
71 citing papers in PubMed, 1 synthesis or guideline pooled it, 240 citations in OpenAlex.
- The coupon collector and the suppressor mutation: estimating the number of compensatory mutations by maximum likelihood.Genetics · 2005Pooled it
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- Simultaneous Affinity Maturation and Liability Removal from Antibodies.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Enhancing monoclonal antibody diversity by integrating bulk sorting and machine learning.Biochemistry and biophysics reports · 2025Article
- Repurposing the bacterial surface display technology for drug delivery.Advanced drug delivery reviews · 2025Review
- Strategies to Screen and Evaluate Brain Targeting Antibodies Using an iPSC-Derived Blood-Brain Barrier Model.Antibodies (Basel, Switzerland) · 2025Article
- Engineering Phages to Fight Multidrug-Resistant Bacteria.Chemical reviews · 2025Review
- A New Bacterial Chassis for Enhanced Surface Display of Recombinant Proteins.Cellular and molecular bioengineering · 2024Article
- Balancing the Affinity and Tumor Cell Binding of a Two-in-One Antibody Simultaneously Targeting EGFR and PD-L1.Antibodies (Basel, Switzerland) · 2024Article
- Genotype-phenotype landscapes for immune-pathogen coevolution.Trends in immunology · 2023Review
- Engineering Ag43 Signal Peptides with Bacterial Display and Selection.Methods and protocols · 2022Article
- Engineering enhanced thermostability into theCurrent research in structural biology · 2022Article
- Predicting antibody affinity changes upon mutations by combining multiple predictors.Scientific reports · 2020Article
- Mutational resilience of antiviral restriction favors primate TRIM5α in host-virus evolutionary arms races.eLife · 2020Article
- Expression, Purification, and Characterization of Anti-International journal of molecular sciences · 2020Article
- Cognizance of Molecular Methods for the Generation of Mutagenic Phage Display Antibody Libraries for Affinity Maturation.International journal of molecular sciences · 2019Review
- Probing pathways of adaptation with continuous evolution.Current opinion in systems biology · 2019Article
- Canine CD117-Specific Antibodies with Diverse Binding Properties Isolated from a Phage Display Library Using Cell-Based Biopanning.Antibodies (Basel, Switzerland) · 2019Article
- Epitope-specific affinity maturation improved stability of potent protease inhibitory antibodies.Biotechnology and bioengineering · 2018Article
11 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Random mutagenesis and selection using phage or cell surface display provides an efficient method for affinity maturation of single chain Fv (scFv) antibodies, thereby improving function in various applications. To investigate the effects of mutation frequency on affinity maturation, error-prone PCR was used to generate libraries containing an average (m) of between 1.7 and 22.5 base substitutions per gene in a high affinity scFv antibody that binds to the cardiac glycoside digoxigenin. The scFv antibody libraries were displayed on Escherichia coli, and mutant populations were analyzed by flow cytometry. At low to moderate mutation frequencies with an average mutation rate of m </= 8, the fraction of clones exhibiting binding to a fluorescently labeled conjugate of digoxigenin decreased exponentially (r(2) = 0.99), but the most highly mutated library (m = 22.5) had significantly more active clones than expected relative to this trend. A library with a low error rate (m = 1.7), one with moderate error rate (m = 3.8), and the one with high error rate (m = 22.5) were screened for high affinity clones under conditions of identical stringency using fluorescence-activated cell sorting. After several rounds of enrichment, each of the three libraries yielded clones with improved affinity for the hapten. The moderate and high error rate libraries gave rise to clones exhibiting the greatest affinity improvement. Taken together, our results indicate that (i) functional clones occur at an unexpectedly high frequency in hypermutated libraries, (ii) gain-of-function mutants are well represented in such libraries, and (iii) the majority of the scFv mutations leading to higher affinity correspond to residues distant from the binding site.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.