We have derived a method for isolating functional mAbs against cell surface receptors that are separatelyeither agonists, antagonists, and partial agonists. We termed the method Directed Ligand Binding (DLB).And in DLB's first iteration (DLB1) we have successfully used it for peptide- and protein ligands. UsingDLB1 these ligands were genetically incorporated into the complementarity determining region (CDR) ofan appropriate phage display scFv library. At its essence DLB is a means to direct or bias the initial bindingof a subtracted (to remove non-specific- and irrelevant binding) phage displayed Ab library toward a targetcell surface receptor binding site by incorporating that receptor's ligand or inhibitor into the Ab library itself.And then rely on the increased binding ability, via the interaction of both an individual mAb's CDRs plusthe covalently attached ligand to transiently stabilize the Ab::receptor complex to withstand the increasingstringency of the phage biopanning washing cycles used to remove any weak binders. In the DLB1method, we genetically encoded the ligand into the Ab's CDR. But that limited us to peptide and proteinligands. We now want to apply the DLB method for the enzymatic conjugation of ligands into anappropriately modified CDR of a complementing library. We refer to this new method as DLB2.0. Thesignificant advantages of the DLB2.0 method are several fold, including: (1) the same library can be usedfor many screens, and (2) the means for testing mAbs derived from DLB2.0 with and without attachedligand is trivial; just omit the ligand attachment step before any screen. The proposed study uses severalGPCR targets as the model and functionality in a cell-based reporter assay as the application. Regardingthe commercial application, we are mainly focused on the method. It is true that the targets we havechosen to develop the method, in some cases, do not have exceptional commercial value. When theDLB2.0 generates appropriate IgGs we intend to partner and collaborate with strong academic, andpharmaceutical- and biotech companies by providing them with the IgG clones and/or purified IgG proteinsso that they can perform the structural and biological studies needed to both better understand and utilizethe mAbs we develop. At Abbratech intend to provide custom services to commercial entities(Pharmaceutical and Biotechnology companies) using the data obtained from this work, including anycollaborative results, as a demonstration of the power of DLB2.0.
Public Health Relevance Statement: "Method for the isolation of antibodies with functional activity against cell surface targets" NARRATIVE We have developed a means to direct or bias the initial binding of a genetically modified phage displayed antibody library toward a target cell surface receptor ligand binding site. In some cases, targeting the site in the method yields antibodies with functional activity. As examples agonistic, antagonists, partial agonists. We now want to apply the method for an enzymatic conjugation of ligands into an appropriately modified antibody of a complementing library. The significant advantages of this new method are several fold, including: (1) the same library can be used for many screens, and (2) the means for testing mAbs derived from the screens is much easier. The proposed study uses several GPCR targets as the model and functionality in a cell-based reporter assay as the application. Regarding the commercial application, we are mainly focused on the method. We intend to partner with strong academic, and pharmaceutical- and biotech companies by providing custom services to them and supply them with the IgG clones and/or purified IgG proteins so that they can perform the structural and biological studies needed to both better understand and utilize the mAbs we develop. Finally, we propose to extend the method beyond GPCRs and start investigating its use for developing functional antibodies against ion channels.
Project Terms: <7S Gamma Globulin>|
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