ArticleJournal of immunological methods2017
A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning.
Article in Journal of immunological methods, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- KREPA6 functions in RNA editing catalytic complex structural organization and gRNA utilization in Trypanosoma brucei.Nucleic acids research · 2026Article
- A Highly Specific Antibody-Based Assay forViruses · 2025Article
- High-throughput strategies for monoclonal antibody screening: advances and challenges.Journal of biological engineering · 2025Review
- Identifying Plasmodium P36 and P52 antigens for coadministration with circumsporozoite protein to enhance vaccine efficacy.NPJ vaccines · 2024Article
- Site-directed neutralizing antibodies targeting structural sites on SARS-CoV-2 spike protein.New biotechnology · 2024Article
- Frequency-potency analysis of IgG+ memory B cells delineates neutralizing antibody responses at single-cell resolution.Cell reports · 2024Article
- A recombinant 9E1 monoclonal antibody binds membrane and soluble channel catfish immunoglobulin M.Fish and shellfish immunology reports · 2023Article
- Biodistribution and function of coupled polymer-DNA origami nanostructures.Scientific reports · 2023Article
- Validation of Ligand Tetramers for the Detection of Antigen-Specific Lymphocytes.Journal of immunology (Baltimore, Md. : 1950) · 2023Article
- Rapid, simple, and effective strategy to produce monoclonal antibodies targeting protein structures using hybridoma technology.Journal of biological engineering · 2023Article
- Antibody-Secreting Cell Isolation from Different Species for Microfluidic Antibody Hit Discovery.Methods in molecular biology (Clifton, N.J.) · 2023Article
- The neutralizing breadth of antibodies targeting diverse conserved epitopes between SARS-CoV and SARS-CoV-2.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Germinal center activity and B cell maturation are associated with protective antibody responses against Plasmodium pre-erythrocytic infection.PLoS pathogens · 2022Article
- Anti-TRAP/SSP2 monoclonal antibodies can inhibit sporozoite infection and may enhance protection of anti-CSP monoclonal antibodies.NPJ vaccines · 2022Article
- Streamlining the Transition From Yeast Surface Display of Antibody Fragment Immune Libraries to the Production as IgG Format in Mammalian Cells.Frontiers in bioengineering and biotechnology · 2022Article
- Protective effects of combining monoclonal antibodies and vaccines against the Plasmodium falciparum circumsporozoite protein.PLoS pathogens · 2021Article
- Antibody interference by a non-neutralizing antibody abrogates humoral protection against Plasmodium yoelii liver stage.Cell reports · 2021Article
- Platelet derived growth factor receptor β (PDGFRβ) is a host receptor for the human malaria parasite adhesin TRAP.Scientific reports · 2021Article
- Targeted Therapy in Cardiovascular Disease: A Precision Therapy Era.Frontiers in pharmacology · 2021Review
- Rapid and sensitive detection of SARS-CoV-2 antibodies by biolayer interferometry.Scientific reports · 2020Article
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Authors and funding
8 authors.
Funding
Abstract
Monoclonal antibody technologies have enabled dramatic advances in immunology, the study of infectious disease, and modern medicine over the past 40years. However, many monoclonal antibody discovery procedures are labor- and time-intensive, low efficiency, and expensive. Here we describe an optimized mAb discovery platform for the rapid and efficient isolation, cloning and characterization of monoclonal antibodies in murine systems. In this platform, antigen-binding splenic B cells from immunized mice are isolated by FACS and cocultured with CD40L positive cells to induce proliferation and mAb production. After 12days of coculture, cell culture supernatants are screened for antigen, and IgG positivity and RNA is isolated for reverse-transcription. Positive-well cDNA is then amplified by PCR and the resulting amplicons can be cloned into ligation-independent expression vectors, which are then used directly to transfect HEK293 cells for recombinant antibody production. After 4days of growth, conditioned medium can be screened using biolayer interferometry for antigen binding and affinity measurements. Using this method, we were able to isolate six unique, functional monoclonal antibodies against an antigen of the human malaria parasite Plasmodium falciparum. Importantly, this method incorporates several important advances that circumvent the need for single-cell PCR, restriction cloning, and large scale protein production, and can be applied to a wide array of protein antigens.
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