ZnO nanomulberry and its significant nonenzymatic signal enhancement for protein microarray

It is very challenging to make a highly sensitive protein microarray because of its lack of a universal signal amplification method like PCR used in DNA microarray. The current strategies to improve the sensitivity mainly rely on a unique nanostructured substrate or enzymatically catalyzed signal amplification, of which the former requires a complicated and time-consuming fabrication process while the latter suffers from high cost and poor stability of enzymes as well as downstream biochemical reactions. In this work, an inexpensive ZnO nanomulberry (NMB) decorated glass slide is investigated as a superior substrate to nonenzymatically amplify the signal of protein microarray for sensitive detection, accomplishing a limit of detection (LOD) of 1 pg mL(-1) and a broad dynamic range of 1 pg mL(-1) to 1 μg mL(-1) to detect an important cancer biomarker, carcinoembryonic antigen (CEA) in 10% human serum. The excellent performance is attributed to ZnO NMB possessing high-density loading of capture antibody and intrinsic enhancement of fluorescence emission. The substrate preparation is simple without using any expensive equipment and complicated technique while offering advantages of low autofluorescence, versatility for various fluorophores, and excellent compatibility with existing microarray fabrication techniques. Thus, a ZnO NMB based protein microarray holds great promise for developing a low cost, sensitive, and high throughput protein assay platform for broad applications in both fundamental research and clinical diagnosis.