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- W2951087716 abstract "Generating recombinant monoclonal antibodies (R-mAbs) from mAb-producing hybridomas offers numerous advantages that increase the effectiveness, reproducibility, and transparent reporting of research. We report here the generation of a novel resource in the form of a library of recombinant R-mAbs validated for neuroscience research. We cloned immunoglobulin G (IgG) variable domains from cryopreserved hybridoma cells and input them into an integrated pipeline for expression and validation of functional R-mAbs. To improve efficiency over standard protocols, we eliminated aberrant Sp2/0-Ag14 hybridoma-derived variable light transcripts using restriction enzyme treatment. Further, we engineered a plasmid backbone that allows for switching of the IgG subclasses without altering target binding specificity to generate R-mAbs useful in simultaneous multiplex labeling experiments not previously possible. The method was also employed to rescue IgG variable sequences and generate functional R-mAbs from a non-viable cryopreserved hybridoma. All R-mAb sequences and plasmids will be archived and disseminated from open source suppliers." @default.
- W2951087716 created "2019-06-27" @default.
- W2951087716 creator A5004778937 @default.
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- W2951087716 date "2019-01-22" @default.
- W2951087716 modified "2023-10-10" @default.
- W2951087716 title "A toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain" @default.
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- W2951087716 doi "https://doi.org/10.7554/elife.43322" @default.
- W2951087716 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6377228" @default.
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- W2951087716 hasPublicationYear "2019" @default.
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