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Updated: Nov 14, 2025

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening
Published on: May 13, 2020
Rapid build-up of high-throughput screening microarrays with biochemistry gradients via light-induced thiol-ene
Hongye Hao1, Junjie Huang, Ping Liu
1MOE Key Laboratory of Macromolecule Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China. jijian@zju.edu.cn.
Abstract:
Microarrays have become extremely powerful experimental tools for high-throughput screening of cell behaviors in multivariate microenvironments. Herein, a microarray-based high-throughput platform with biochemistry gradients was developed using poly(limonene carbonate) (PLimC) as a substrate through thiol-ene click chemistry. ATR-IR, XPS, Raman spectrum, and water contact angle results demonstrated that the sulfhydryl molecules, including PEG (polyethylene glycol) and RGD (arginine-glycine-aspartic acid) peptide, could be grafted onto PLimC substrates, while the grafting density could be well controlled by regulating the intensity of UV irradiation. Then, microarrays with a gradient of RGD grafting density were fabricated by using UV irradiation patterned by a photomask and a gradient light filter. Adhesion experiments of smooth muscle cells and 3T3-L1 mouse embryonic fibroblast cells proved that the cell behaviors were highly determined by the RGD density. This platform puts forward a facile, high-throughput method to study the effect of biochemical signal density on cell behaviors.

