Protein immobilization on plasma-polymerized ethylenediamine-coated glass slides

Abstract

For protein chip construction, protein immobilization on the surface of the glass slide is essential. It was previously reported that glass slides are embedded with chemicals that contain primary amines and aldehydes for protein immobilization. We fabricated a plasma-polymerized ethylenediamine (PPEDA)-coated slide that exposed primary amines. For the plasma polymer deposition on the glass slide, the inductively coupled plasma (ICP) power was found to be a critical factor in sustaining a high density of amine on the surface of the PPEDA films. We prepared PPEDA-coated slides at three different ICP powers (3, 30, or 70 W). In the slide that was prepared at a low ICP power (3 W), we detected a high density of primary amine. Therefore, the fluorescein isothiocyanate-conjugated immunoglobulin G (IgG) was highly immobilized to the PPEDA-coated slide that was prepared at the low ICP power. For protein immobilization, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) was used as a cross-linker. The immobilization of the protein to the PPEDA-coated slide was carried out by consecutive incubations with 1 mg/ml EDC for 5 min and 0.1 mg/ml IgG for 1 h. This efficiently produced the functionally active protein-immobilized slide. Therefore, this work shows that the plasma technique can be applied to produce a high-quality glass surface for the immobilization of proteins and other materials.