A microfluidic technique to estimate antigen expression on particles.

Abstract

Antigen expression is an important biomarker for cell analysis and disease diagnosis. Traditionally, antigen expression is measured using a flow cytometer which, due to its cost and labor intensive sample preparation, is unsuitable to be used at the point-of-care. Therefore, an automatic, miniaturized assay which can measure antigen expression in the patient could aid in making crucial clinical decisions rapidly. Such a device would also expand the use of such an assay in basic research in biology. In this paper, we present a microfluidic device that can be used to measure antigen expression on cells. We demonstrate our approach using biotin-neutravidin as the binding pair using experimental and computational approaches. We flow beads with varying biotin surface densities (mr) through a polydimethylsiloxane channel with cylindrical pillars functionalized with neutravidin. We analyze how shear stress and collision angle, the angle at which the beads collide with the pillars, affect the angular location of beads captured on the pillars. We also find that the fraction of captured beads as a function of distance (γ) in the channel is affected by mr. Using γ, we derive the probability of capture per collision with the pillar (ε). We show that ε is linearly related to mr, which is analogous to the expression level of proteins on cell surfaces. Although demonstrated with beads, this assay can next be expanded with cells, thus paving the way for a rapid antigen expression test.