title: Poly(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) Brushes as Peptide/Protein Microarray Substrate for Improving Protein Binding and Functionality.
abstract: We developed a three-dimensional (3D) polymer-brush substrate for protein and peptide microarray fabrication, and this substrate was facilely prepared by copolymerization of glycidyl methacrylate (GMA) and 2-hydroxyethyl methacrylate (HEMA) monomers via surface-initiated atom transfer radical polymerization (SI-ATRP) on a glass slide. The performance of obtained poly(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) (P(GMA-HEMA)) brush substrate was assessed by binding of human IgG with rabbit antihuman IgG antibodies on a protein microarray and by the determination of matrix metalloproteinase (MMP) activities on a peptide microarray. The P(GMA-HEMA) brush substrate exhibited higher immobilization capacities for proteins and peptides than those of a two-dimensional (2D) planar epoxy slide. Furthermore, the sensitivity of the P(GMA-HEMA) brush-based microarray on rabbit antihuman IgG antibody detection was much higher than that of its 2D counterpart. The enzyme activities of MMPs were determined specifically with a low detection limit of 6.0 pg mL(-1) for MMP-2 and 5.7 pg mL(-1) for MMP-9. By taking advantage of the biocompatibility of PHEMA, the P(GMA-HEMA) brush-based peptide microarray was also employed to evaluate the secretion of MMP-2 and MMP-9 by cells cultured off the chip or directly on the chip, and satisfactory results were obtained.
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022, P. R. China.|University of Chinese Academy of Sciences , Beijing 100049, P. R. China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Lei
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022, P. R. China.|University of Chinese Academy of Sciences , Beijing 100049, P. R. China.
authors.forename: Jiaxue
authors.initials: J
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022, P. R. China.
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022, P. R. China.
authors.forename: Dianjun
authors.initials: D
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022, P. R. China.
authors.forename: Zhenxin
authors.initials: Z
authors.lastname: Wang
mesh_terms: D004852:Epoxy Compounds; D006801:Humans; D008689:Methacrylates; D010455:Peptides / Q000032:analysis*; D040081:Protein Array Analysis; D011485:Protein Binding
chemical_list: D004852:Epoxy Compounds; D008689:Methacrylates; D010455:Peptides; C005044:hydroxyethyl methacrylate; C007870:glycidyl methacrylate
keywords: binding of human IgG with rabbit antihuman IgG antibody; matrix metalloproteinase; peptide microarray; poly(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) P(GMA-HEMA); polymer-brush substrate; protein microarray