ArticlePloS one2014
Comprehensive evaluation and optimization of amplicon library preparation methods for high-throughput antibody sequencing.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- The Evolution of Artificial Intelligence in Antibody Design: From Structure-Based Engineering to Generative Models.Antibodies (Basel, Switzerland) · 2026Review
- Dynamic thresholding and tissue dissociation optimization for CITE-seq identifies differential surface protein abundance in metastatic melanoma.Communications biology · 2023Article
- Phenotypic determinism and stochasticity in antibody repertoires of clonally expanded plasma cells.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Individualized VDJ recombination predisposes the available Ig sequence space.Genome research · 2021Article
- Immunoglobulin germline gene variation and its impact on human disease.Genes and immunity · 2021Review
- Optimization of therapeutic antibodies by predicting antigen specificity from antibody sequence via deep learning.Nature biomedical engineering · 2021Article
- A single donor is sufficient to produce a highly functional in vitro antibody library.Communications biology · 2021Article
- CRIS: complete reconstruction of immunoglobulinBioinformatics advances · 2021Article
- Article
- Uncoupling of invasive bacterial mucosal immunogenicity from pathogenicity.Nature communications · 2020Article
- Chronic Viral Infection Promotes Efficient Germinal Center B Cell Responses.Cell reports · 2020Article
- Large-scale network analysis reveals the sequence space architecture of antibody repertoires.Nature communications · 2019Article
- High-Quality Library Preparation for NGS-Based Immunoglobulin Germline Gene Inference and Repertoire Expression Analysis.Frontiers in immunology · 2019Article
- Molecular Landscape of Anti-Drug Antibodies Reveals the Mechanism of the Immune Response Following Treatment With TNFα Antagonists.Frontiers in immunology · 2019Article
- Metabolic and Transcriptional Modules Independently Diversify Plasma Cell Lifespan and Function.Cell reports · 2018Article
- High-throughput antibody engineering in mammalian cells by CRISPR/Cas9-mediated homology-directed mutagenesis.Nucleic acids research · 2018Article
- Profiling of the TCRβ repertoire in non-model species using high-throughput sequencing.Scientific reports · 2018Article
- Effects of spaceflight on the immunoglobulin repertoire of unimmunized C57BL/6 mice.Life sciences in space research · 2018Article
- Synthetic Standards Combined With Error and Bias Correction Improve the Accuracy and Quantitative Resolution of Antibody Repertoire Sequencing in Human Naïve and Memory B Cells.Frontiers in immunology · 2018Article
- Production of individualized V gene databases reveals high levels of immunoglobulin genetic diversity.Nature communications · 2016Article
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-throughput sequencing (HTS) of antibody repertoire libraries has become a powerful tool in the field of systems immunology. However, numerous sources of bias in HTS workflows may affect the obtained antibody repertoire data. A crucial step in antibody library preparation is the addition of short platform-specific nucleotide adapter sequences. As of yet, the impact of the method of adapter addition on experimental library preparation and the resulting antibody repertoire HTS datasets has not been thoroughly investigated. Therefore, we compared three standard library preparation methods by performing Illumina HTS on antibody variable heavy genes from murine antibody-secreting cells. Clonal overlap and rank statistics demonstrated that the investigated methods produced equivalent HTS datasets. PCR-based methods were experimentally superior to ligation with respect to speed, efficiency, and practicality. Finally, using a two-step PCR based method we established a protocol for antibody repertoire library generation, beginning from inputs as low as 1 ng of total RNA. In summary, this study represents a major advance towards a standardized experimental framework for antibody HTS, thus opening up the potential for systems-based, cross-experiment meta-analyses of antibody repertoires.
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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.