Development of Dysferlin Affinity Binders

Silvère van der Maarel, PhD

Leiden University Medical Center (Leiden The, Netherlands)

Dr. van der Maarel is a professor in the Department of Human Genetics at Leiden University Medical Center (Leiden, The Netherlands).

Past Projects

Objective: 
Generation of a comprehensive set of dysferlin-specific affinity binders

Two major isoforms of DYSF are described that differ in their first exon, which in both cases encode part of the first C2 domain. It is not known to what extent these isoforms are functionally different. We plan to generate antibodies against the two alternate isoforms, to allow studies of their functions. We are also generating and characterizing antibodies against dysferlin C2 domains, the DysF domain, the extracellular C-terminal domain (~12 amino acids); endogenous dysferlin (including yet unknown post-translational modifications); and the interaction partners of dysferlin. We aim to implement the selected binders in intra-body vectors to get insight into dysferlin function by: 1) monitoring the intracellular localization of endogenous and recombinant dysferlin in various conditions such as Ca2+ depletion, membrane injury, and shRNA knockdown of interaction partners; and 2) modulation of mutant dysferlin localization following Ca++ depletion and membrane injury. We will also assess changes in mutant dysferlin following knockdown of known binding proteins as described above.