ArticlePloS one2015
Effective Optimization of Antibody Affinity by Phage Display Integrated with High-Throughput DNA Synthesis and Sequencing Technologies.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 53 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
53 citing papers in PubMed, 90 citations in OpenAlex.
- Article
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- Review
- Therapeutic Antibodies for Infectious Diseases: Recent Past, Present, and Future.Biochemistry · 2025Review
- The characterization of variable new antigen receptors targeting FAP isolated from a novel immunized library.Communications biology · 2025Article
- FASTIA: A rapid and accessible platform for protein variant interaction analysis demonstrated with a single-domain antibody.Protein science : a publication of the Protein Society · 2025Article
- Development of FAP-targeted theranostics discovered by next-generation sequencing-augmented mining of a novel immunized VNAR library.bioRxiv : the preprint server for biology · 2025Article
- Beyond Single Clones: High-Throughput Sequencing in Antibody Discovery.Cold Spring Harbor protocols · 2025Article
- Optimizing Yeast Homologous Recombination for Splicing Large Coronavirus Genome Fragments.International journal of molecular sciences · 2024Article
- Optimization of PCA Error Correction Conditions to Improve Efficiency of Virus Genome De Novo Synthesis.International journal of molecular sciences · 2024Article
- Bacterial cell surface characterization by phage display coupled to high-throughput sequencing.Nature communications · 2024Article
- Development of a new affinity maturation protocol for the construction of an internalizing anti-nucleolin antibody library.Scientific reports · 2024Article
- Balancing the Affinity and Tumor Cell Binding of a Two-in-One Antibody Simultaneously Targeting EGFR and PD-L1.Antibodies (Basel, Switzerland) · 2024Article
- The rise of big data: deep sequencing-driven computational methods are transforming the landscape of synthetic antibody design.Journal of biomedical science · 2024Review
- Influencing factors and solution strategies of chimeric antigen receptor T-cell therapy (CAR-T) cell immunotherapy.Oncology research · 2024Review
- Review on Monoclonal Antibodies (mAbs) as a Therapeutic Approach for Type 1 Diabetes.Current diabetes reviews · 2024Review
- Design, optimization, production and activity testing of recombinant immunotoxins expressed in plants and plant cells for the treatment of monocytic leukemia.Bioengineered · 2023Article
- Position-Specific Enrichment Ratio Matrix scores predict antibody variant properties from deep sequencing data.Bioinformatics (Oxford, England) · 2023Article
- AI/ML combined with next-generation sequencing of VHH immune repertoires enables the rapid identification ofFrontiers in molecular biosciences · 2023Article
Corrections and comments
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Authors and funding
10 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phage display technology has been widely used for antibody affinity maturation for decades. The limited library sequence diversity together with excessive redundancy and labour-consuming procedure for candidate identification are two major obstacles to widespread adoption of this technology. We hereby describe a novel library generation and screening approach to address the problems. The approach started with the targeted diversification of multiple complementarity determining regions (CDRs) of a humanized anti-ErbB2 antibody, HuA21, with a small perturbation mutagenesis strategy. A combination of three degenerate codons, NWG, NWC, and NSG, were chosen for amino acid saturation mutagenesis without introducing cysteine and stop residues. In total, 7,749 degenerate oligonucleotides were synthesized on two microchips and released to construct five single-chain antibody fragment (scFv) gene libraries with 4 x 10(6) DNA sequences. Deep sequencing of the unselected and selected phage libraries using the Illumina platform allowed for an in-depth evaluation of the enrichment landscapes in CDR sequences and amino acid substitutions. Potent candidates were identified according to their high frequencies using NGS analysis, by-passing the need for the primary screening of target-binding clones. Furthermore, a subsequent library by recombination of the 10 most abundant variants from four CDRs was constructed and screened, and a mutant with 158-fold increased affinity (Kd = 25.5 pM) was obtained. These results suggest the potential application of the developed methodology for optimizing the binding properties of other antibodies and biomolecules.
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Registered trials
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.