Micropatterned antibody-terminated nanocomposites (MANs) fabricated using layer-by-layer lift-off (LBL-LO) technique

Patterned functional biomolecular surfaces have been created using a wide range of fabrication methods for applications in cell-based studies, with the most common methods based on photolithography or soft lithography. In this study, a simple fabrication method combining photolithography, layer-by-layer (LbL) self-assembly, and photoresist lift-off (LO) was used to create spatially defined nanocomposite micropatterns of antibodies on polyelectrolyte-protein multilayer beds. The two components of the micropatterned surfaces consisted of an anti-CD44 rat monoclonal antibody atop a vitronectin-coated multilayer film and an anti-rat osteopontin (MPIIIB10(1)) anitibody atop a fibronectin-coated multilayer film. The micropatterned antibody-terminated nanocomposites (MANs) were characterized using bright field microscopy, surface profilometry, and atomic force microscopy. The results demonstrate that LbL-LO method can be successfully used to create patterns of proteins and antibodies in precise spatial arrangements. The MAN surfaces can be utilized in many bionanotechnology applications, including patterned cell cocultures or tricultures, cytotoxicity testing, drug screening, and biosensing. In addition, the general fabrication approach used in this study is applicable to other antibodies and other ligands.