Paper detail

Highly efficient intracellular chromobody delivery by mesoporous silica nanoparticles for antigen targeting and visualization in real time

Chromobodies have recently drawn great attention as bioimaging nanotools. They offer antigen binding specificity and affinity comparable to conventional antibodies, but much smaller size and higher stability. Importantly, chromobodies can be used in live cell imaging for highly specific spatio-temporal visualization of cellular processes. To date, functional application of chromobodies requires lengthy genetic manipulation of the target cell. Here, we developed multifunctional large-pore mesoporous silica nanoparticles (MSNs) as nanocarriers to directly transport chromobodies into living cells for antigen-visualization in real time. The multifunctional large-pore MSNs feature high loading capacity for chromobodies, and are efficiently taken up by cells. By functionalizing the internal MSN surface with nitrilotriacetic acid-metal ion complexes, we could control the release of His6-tagged chromobodies from MSNs in acidified endosomes. When chromobodies escape from the endosomes through the proton sponge effect generated by their built-in His6-tags, co-localization of signals from fluorescent endogenous antigen and organic dye-labeled chromobodies can be detected. Hence, by combining the two nanotools, chromobodies and MSNs, we established a new powerful approach for chromobody applications in living cells.