Nanometer-scale antibody patterning for directed eosinophil cell immobilization and stimulation

Antibodies (Ab) are patterned at nanoscale precision for the precise immobilization and stimulation of immune cells. We demonstrate that the antigen bovine serum albumin (BSA) can be patterned on silicon using a photolithographically patterned polymer lift-off technique. The nanoscale pattern is realized as the polymer is mechanically peeled away in one contiguous piece in aqueous solution. Anti-BSA Ab are bound specifically to BSA to create a pattern of oriented Ab that provides a surface for eosinophil immobilization and degranulation. The patterns ranged from 0.36 μm² to 4,489 μm², appropriate dimensions for the 10-14 μm diameter eosinophil cells. This method provides a new technique for immobilizing cells onto nano and micrometer scale patterns for analyzing cellular biochemical cascade events such as degranulation and studying cellular morphological changes in response to defined nanoscale antigenic stimulus