There is a continual need for high-quality binding reagents that report on the structural conformation, localization, spatiotemporal dynamics and interaction factors of cellular components. Single-domain antibody fragments – referred to as nanobodies - derived from heavy-chain-only antibodies of camelids have emerged as attractive alternatives to conventional antibodies for multiple applications in biomedical research. Nanobodies provide an outstanding tool for structural analysis, imaging and proteome analysis as they are small (~ 15 kDa), highly soluble and stable, and can be targeted-modified according to the needs of the intended application.

Our research is focused on:

(a) the development, functionalization and application of novel nanobodies for protein purification, protein-protein interaction analysis, crystallization studies, mass spectrometry approaches and super-resolution microscopy

(b) the generation and validation of intracellular biosensors (chromobodies) to monitor components and processes in living cells using fluorescence microscopy including high-content imaging

Covalent coupling of nanobodies via termini-specific side chains or by the use of transpeptidases ensures a site-directed labeling with organic dyes or oriented immobilization on various matrices like agarose, microspheres or chip surfaces yielding a high binding capacity accompanied with minimal steric hindrances and low non-specific background.

For in cellulo studies we make use of intracellularly functional nanobodies (chromobodies) to monitor the spatiotemporal dynamics and the abundance of endogenous components in living cells. In combination with high-throughput microscopy and automated image analysis we apply chromobodies as intracellular biosensors for phenotypic screening and high-content imaging (HCI) in real time. To date chromobody-based cellular models have been established to monitor cell cycle, signal transduction or to trace epithelial mesenchymal transition upon compound treatment within living cells.

Based on a long standing expertise in recombinant antibody technologies we take advantage of the extraordinary properties of nano- and chromobodies to generate versatile binding molecules offering a unique opportunity to combine biochemical, structural and functional analyses of multiple cellular targets in flexible settings.