A General Method for Patterning Gradients of Biomolecules on Surfaces Using Microfluidic Networks

Abstract

This report outlines a general method for the fabrication of immobilized gradients of biomolecules on surfaces. This method utilizes a microfluidic network that generates a gradient of avidin in solution and immobilizes this protein on the surface of glass or poly(dimethylsiloxane) by physical adsorption. The immobilized gradient of avidin is then translated into gradients of biotinylated ligands (e.g., small molecules, oligomers of DNA, polysaccharides) using the specific interaction between biotin and avidin. This method can also generate immobilized gradients of certain proteins and artificial polymers by a direct transfer of gradients from solution onto the surface. The major advantage of this method is that almost any type of molecule can, in principle, be immobilized in a well-defined surface gradient of arbitrary shape with dimensions of a few micrometers to a few centimeters. It is possible to tailor the precise shapes of gradients on surfaces from gradients in solution, either kinetically or competitively. Kinetic methods rely on controlling the time that the surface is exposed to the gradient in solution: when a single protein adsorbs from solution, the amount that adsorbs depends both on its concentration in solution and on the time allowed for adsorption. Competitive methods rely on exposure of the surface to a complementary gradient of two proteins in solution (In these experiments, the sum of the concentrations of the proteins in solution is independent of positions although the concentration of each, individually, depends on the position. In this procedure, the relative amount of each protein, at saturation on the surface, depends only on its concentration.).