G.P.7 05 Reliable and controllable antibody fragment selections from Camelid non-immune libraries

Muscular dystrophies comprise a heterogeneous group of muscle disorders often caused by mutations in genes encoding proteins that participate in large protein complexes. To fully understand the function of these proteins and the consequences of their dysfunction, preferably these proteins should be studied in their natural context with a combination of biochemical and microscopic techniques using specific antibodies. Therefore, we designed a fast, reliable and controllable selection and screening method of single-domain antibody fragments (VHH) from a Camelid non-immune library. We isolated VHH for five muscle disease related proteins; emerin, actin, tropomyosin-1, dysferlin, and nuclear poly(A)-binding protein. We show that selected antibody fragments are functional in various immunological techniques and prove useful in diagnostic applications. Our selection strategy is amenable to automation and to the establishment of proteomics platforms. It opens the way to quickly and cost-effectively obtain multiple antibody fragments for many antigens that can detect changes in their localization, level, and modification as well as subtle changes in supramolecular structures, which often associate with disease.