Investigation of non-nucleophilic additives for reduction of morphological anomalies in protein arrays

摘要

Uniform spot morphology is of critical importance in the fabrication and successful use of protein arrays, and solution additives are often needed in order to ensure good spot quality. While hydroxyl-bearing molecules such as glycerol have found wide usage, in our experience these reduce the efficiency of probe immobilization (particularly in the context of aldehyde-terminated surfaces). Here, we report a series of non-nucleophilic molecules that can be used as additives in order to improve spot homogeneity in protein arrays. Arrayed Imaging Reflectometry, a label-free optical biosensing technique, has been used along with spectroscopic ellipsometry to test the spot homogeneity, antibody immobilization efficiency, and activity of anti-human IgG arrays prepared with these non-nucleophilic additives on glutaraldehyde surfaces. It has been determined that 0.1% v/v 12-crown-4 has the optimum performance in MPBS buffer.