ArticlePloS one2019
A simplified workflow for monoclonal antibody sequencing.
Article in PloS one, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers.
What it found
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Who cites it
52 citing papers in PubMed, 85 citations in OpenAlex.
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- A single-chain derivative of an integrin-activating antibody potentiates organoid growth in Matrigel and collagen hydrogels.Nature biotechnology · 2026Article
- Discovery, Structural Characterization, and Preclinical Evaluation of Monoclonal Antibodies against Xylazine Poisoning.ACS pharmacology & translational science · 2026Article
- Novel recombinant VH43 antibody fragment potentiates the antibacterial activity of cephalosporins against Enterococcus faecium.Archives of microbiology · 2026Article
- Idiotype-specific CD4Nature communications · 2026Article
- Transgenic mouse-derived human monoclonal antibodies targeting EBV gp350 and gp42 provide basis for therapeutic development.Cell reports. Medicine · 2026Article
- Discovery of potent cross-neutralizing antibodies against Marburg and Ravn viruses with therapeutic potential.Npj viruses · 2025Article
- Sources of essential lipids for Mycoplasma pneumoniae via P116 to target liver and atherosclerotic lesions.Nature communications · 2025Article
- Expression of porcine-derived IgA against PEDV and use of Alg-CS gel to protect the IgA from degradation in simulated gastric fluid.Archives of virology · 2025Article
- A Novel FLI1 Monoclonal Antibody Which Recognizes EWS::FLI1 with High Affinity Is Useful for Detecting Ewing Sarcoma.Antibodies (Basel, Switzerland) · 2025Article
- Interdomain disulfide bonds of rabbit kappa light chain allotypes influence mouse-rabbit chimeric antibody performance.bioRxiv : the preprint server for biology · 2025Article
- Cost-Effective Method for Full-Length Sequencing of Monoclonal Antibodies from Hybridoma Cells.Antibodies (Basel, Switzerland) · 2025Article
- Open-source antibodies as a path to enhanced research reproducibility and transparency.New biotechnology · 2025Article
- clonevdjseq: A workflow and bioinformatics management system for sequencing, archiving, and analysis of VDJ sequences from clonal libraries.BMC bioinformatics · 2025Article
- Monoclonal antibody neutralizesInfection and immunity · 2025Article
- Structure of endogenous Pfs230:Pfs48/45 in complex with potent malaria transmission-blocking antibodies.bioRxiv : the preprint server for biology · 2025Article
- ZP4: A novel target for CAR-T cell therapy in triple negative breast cancer.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Article
- A type-specific B-cell epitope at the apex of outer surface protein C (OspC) of the Lyme disease spirochete,Microbiology spectrum · 2025Article
- Discovery of three novel neutralizing antibody epitopes on the human astrovirus capsid spike and mechanistic insights into virus neutralization.Journal of virology · 2025Article
- Identification of specific neutralizing antibodies for highly pathogenic avian influenza H5 2.3.4.4b clades to facilitate vaccine design and therapeutics.Emerging microbes & infections · 2024Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
Abstract
The diversity of antibody variable regions makes cDNA sequencing challenging, and conventional monoclonal antibody cDNA amplification requires the use of degenerate primers. Here, we describe a simplified workflow for amplification of IgG antibody variable regions from hybridoma RNA by a specialized RT-PCR followed by Sanger sequencing. We perform three separate reactions for each hybridoma: one each for kappa, lambda, and heavy chain transcripts. We prime reverse transcription with a primer specific to the respective constant region and use a template-switch oligonucleotide, which creates a custom sequence at the 5' end of the antibody cDNA. This template-switching circumvents the issue of low sequence homology and the need for degenerate primers. Instead, subsequent PCR amplification of the antibody cDNA molecules requires only two primers: one primer specific for the template-switch oligonucleotide sequence and a nested primer to the respective constant region. We successfully sequenced the variable regions of five mouse monoclonal IgG antibodies using this method, which enabled us to design chimeric mouse/human antibody expression plasmids for recombinant antibody production in mammalian cell culture expression systems. All five recombinant antibodies bind their respective antigens with high affinity, confirming that the amino acid sequences determined by our method are correct and demonstrating the high success rate of our method. Furthermore, we also designed RT-PCR primers and amplified the variable regions from RNA of cells transfected with chimeric mouse/human antibody expression plasmids, showing that our approach is also applicable to IgG antibodies of human origin. Our monoclonal antibody sequencing method is highly accurate, user-friendly, and very cost-effective.
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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.