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• W3205510020 abstract "The identification of antibody variable regions in the heavy (V H ) and light (V L ) chains from hybridomas is necessary for the production of recombinant, sequence-defined monoclonal antibodies (mAbs) and antibody derivatives. This process has received renewed attention in light of recent reports of hybridomas having unintended specificities due to the production of non-antigen specific heavy and/or light chains for the intended antigen. Here we report a surprising finding and potential pitfall in variable domain sequencing of an anti-human CD63 hybridoma. We amplified multiple V L genes from the hybridoma cDNA, including the well-known aberrant Sp2/0 myeloma V K and a unique, full-length V L . After finding that the unique V L failed to yield a functional antibody, we discovered an additional full-length sequence with surprising similarity (~95% sequence identify) to the non-translated myeloma kappa chain but with a correction of its key frameshift mutation. Expression of the recombinant mAb confirmed that this highly homologous sequence is the antigen-specific light chain. Our results highlight the complexity of PCR-based cloning of antibody genes and strategies useful for identification of correct sequences." @default.
• W3205510020 created "2021-10-25" @default.
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• W3205510020 date "2021-10-11" @default.
• W3205510020 modified "2023-10-13" @default.
• W3205510020 title "A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain" @default.
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• W3205510020 doi "https://doi.org/10.1371/journal.pone.0252558" @default.
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