Neuronal Cell Patterning on Covalently Bound Protein Patterns by Micro-Contact Printing Techniques and the Functioning of Proteins Bound on Silane Monolayers

Micro-patterning of neuronal cells in vitro is a critical step for studies in the fundamental biology of neuron-neuron and neuron-surface interactions. The culturing of neuronal cells on patterned self-assembled monolayers (SAMs) in some cases require further chemical modifications of the SAM surfaces to induce cell adhesion and promote neurite outgrowth. In these cases it would be important to select a negative surface modifying agent such as an extracellular matrix protein like tenascin-C and a specific functional protein like an antibody that interacts with neuronal cell adhesion molecules to support cell adhesion on specially designed surface patterns. The protein modified surfaces could then be used to arrange cells in specific patterns and control cell growth because of a specific protein function such as an inhibition of cell outgrowth. Protein immobilization on solid substrates could then be used as support layers for biosensors such as neuronal cell-based sensors, neuronal networks, biomedical devices, bioprocessing, bioassays, separations, and synthesis. There is an increasing need in the field of biosensors to immobilize a functional protein on silica surfaces. Antibody immobilization on a silica surface can be easily accomplished by direct adsorption. However, this method results in partial denaturation of the protein, as well as an unstable attachment. Consequently, partial loss of protein can occur. Covalent attachments of functional proteins are one solution and are important when a coated substrate is subjected to a flowing solution or exposed for a long period of time in solution. Research indicates that covalent immobilizations of proteins can preserve their functions for up to two years. The original document contains color images.