Capture and display of antibodies secreted by hybridoma cells enables fluorescent on-cell screening.
MABS(2019)
摘要
Hybridoma methods for monoclonal antibody (mAb) cloning are a mainstay of biomedical research, but they are hindered by the need to maintain hybridomas in oligoclonal pools during antibody screening. Here, we describe a system in which hybridomas specifically capture and display the mAbs they secrete: On-Cell mAb Screening (OCMS (TM)). In OCMS (TM), mAbs displayed on the cell surface can be rapidly assayed for expression level and binding specificity using fluorescent antigens with high-content (image-based) methods or flow cytometry. OCMS (TM) demonstrated specific mAb binding to poliovirus and rabies virus by forming a cell surface IgG "cap", as a universal assay for anti-viral mAbs. We produced and characterized OCMS (TM)-enabled hybridomas secreting mAbs that neutralize poliovirus and used fluorescence microscopy to identify and clone a human mAb specific for the human N-methyl-D-aspartate receptor. Lastly, we used OCMS (TM) to assess expression and antigen binding of a recombinant mAb produced in 293T cells. As a novel method to physically associate mAbs with the hybridomas that secrete them, OCMS (TM) overcomes a central challenge to hybridoma mAb screening and offers new paradigms for mAb discovery and production.
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关键词
Monoclonal antibodies,human antibodies,hybridoma,high throughput screening,poliovirus,anti-viral antibodies,NMDA receptor,OCMS (TM)
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