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.

Delamination Kinetics of Thin Film Poly(acrylate) Model Coatings Prepared by Surface Initiated Atom Transfer Radical Polymerization on Iron

“…39 The small peak present at 1731 cm −1 originates from the C=O group present in the head of the molecule. 39 The C-H stretching vibrations of the alkyl groups appeared at 2923 and 2965 cm −1 . OH stretching modes of strongly hydrogen bound water or hydroxyl groups were present, centered at around 3200 cm −1 .…”

Biosensors and bioassays for determination of matrix metalloproteinases: state of the art and recent advances

“…Besides the above described assays/biosensors, some other techniques have also been applied for quantifying MMPs, such as mass-spectrometry (MS)-based proteomic techniques, 179-182 array/microarray-based assays, [183][184][185][186][187][188] and nuclear imaging approaches. [189][190][191] MS and liquid chromatography-tandem MS (LC-MS/MS) have evolved as promising techniques for qualitative and quantitative analysis of proteins via identification of the unique mass signature of the target.…”

In Pursuit of Zero 2.0: Recent Developments in Nonfouling Polymer Brushes for Immunoassays

“…Lei et al synthesized P(GMA-HEMA) brushes as a substrate for protein and peptide microarrays for the detection of rabbit antihuman IgG Abs and matrix metalloproteinases (MMP) activity. [90] While detection of rabbit anti-human IgG Abs was used as a proof-ofconcept study for the platform, MMP activity is related to several pathological processes [91] and is thus potentially important clinically. MMP activity was assessed by immobilizing biotinylated peptide sequences (subsequentially labeled with avidin-Cy3) that can be cleaved by specific MMP enzymes.…”