On-chip microscale isoelectric focusing enhances protein detection limit.

Abstract

Enhancing the detection limit in protein analysis is essential for a wide range of biomedical applications. In typical fluorescent protein assays, this limit is constrained by the detection capacity of the photon detector. Here, we develop an approach that significantly enhances the protein detection threshold by using microscale isoelectric focusing implemented directly at the detection site on a protein sensor chip. We demonstrate that by electrically generating a localized pH environment within a radius of ∼60 μm, protein molecules can be concentrated within this range and be detected at levels over four times lower than those achieved by measurements without on-chip isoelectric focusing. We find that this detection-limit enhancement results from a dual effect: the concentrating of the protein molecules and a reduction in the diffusion-induced fluctuation. Our approach offers a simple, yet highly effective ultra-low-power all-electronic solution for substantially improving protein analysis detection limits for diverse applications, including healthcare, clinical diagnostics, and therapeutics.