Abstract
Monoclonal antibodies (mAbs) can be potent and highly specific therapeutics, diagnostics and research reagents. Nonetheless, mAb discovery using current in vivo or in vitro approaches can be costly and time-consuming, with no guarantee of success. We have established a platform for rapid discovery and optimization of mAbs ex vivo. This DTLacO platform derives from a chicken B cell line that has been engineered to enable rapid selection and seamless maturation of high affinity mAbs. We have validated the DTLacO platform by generation of high affinity and specific mAbs to five cell surface targets, the receptor tyrosine kinases VEGFR2 and TIE2, the glycoprotein TROP2, the small TNF receptor family member FN14, and the G protein-coupled receptor FZD10. mAb discovery is rapid and humanization is straightforward, establishing the utility of the DTLacO platform for identification of mAbs for therapeutic and other applications.
Highlights
Monoclonal antibodies are well-established as therapeutics, diagnostics, and reagents for research, but their use is currently limited by the difficulties and costs associated with identifying mAbs with the required affinity and specificity
We have demonstrated generation of high affinity mAbs against six targets, including the model antigen, streptavidin (SAv), and five cell surface antigens, the receptor tyrosine kinases VEGFR2 and TIE2, the glycoprotein TROP2, the TNF receptor family member FN14, and the G proteincoupled receptor FZD10
The DTLacO mAb discovery platform The DTLacO platform for rapid mAb selection and optimization was developed by engineering DT40 cells to put diversification under control of the powerful E. coli lactose operator (LacO)/LacI regulatory network
Summary
Monoclonal antibodies (mAbs) are well-established as therapeutics, diagnostics, and reagents for research, but their use is currently limited by the difficulties and costs associated with identifying mAbs with the required affinity and specificity. Many key therapeutic targets are cell surface proteins, which present particular challenges to mAb development because their physiologically active conformations are not readily recapitulated by purified proteins or membrane preparations used for immunization to elicit specific antibodies. This includes some especially high value targets, such as cytokine receptors and G protein-coupled receptors. In vitro approaches rely on screening massive numbers of synthetic single-chain antibodies, typically displayed on phage [3,4] These antibodies are expressed by cloned genes that encode linked VH and VL regions derived from an immune repertoire, often from a convalescent individual [5,6]. Success in the end depends on the quality of the starting libraries and their sources, and not all single-chain antibodies can be readily converted to natural antibodies for practical applications
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