Porous Protein‐Based Nanoparticle Hydrogel for Protein Chips with Improved Sensitivity

Abstract

We have discovered a novel method to prepare a protein‐based hydrogel, that is, a ‘three‐dimensional nanostructured protein hydrogel’ (3D NPH), which is composed of loosely inter‐connected protein–polymer hybrid nanoparticles. The 3D NPH can be easily prepared by spotting a protein/polymer mixture on a substrate. Surprisingly, gold nanoparticles carrying protein molecules easily diffuse into the 3D NPH through pores and spaces. We have shown that the protein chip made by our 3D NPH method has tremendously improved sensitivity in detecting protein–protein interactions compared with that by direct protein immobilization methods.

Activated Ester Containing Polymers: Opportunities and Challenges for the Design of Functional Macromolecules

“…The technique required spotting a mixture of protein IgG and a copolymer of poly­(NAM- co -NHSA) on a Au surface functionalized with NHS-ester motifs. Covalent binding of the amine functionality of the protein with NHS-ester of the polymer and/or substrate resulted in highly porous cross-linked networks . Photopolymerization of NHSA, acrylamide, and a cross-linker on top of 2D polystyrene particle array followed by selective removal of PS beads resulted in 2D hole array on hydrogels .…”

Soft-Nanocomposites of Nanoparticles and Nanocarbons with Supramolecular and Polymer Gels and Their Applications

“…Tanaka et al assembled three-dimensional nanostructured protein-based hydrogel composed of "loosely" interconnected protein− polymer hybrid nanoparticles using different proteins (streptavidin, protein A, and Mouse IgG) and a polymer poly[(Nacryloylmorpholine)-co-(N-acryloxysuccinimide)]. 163 The protein hydrogel has been accomplished by spotting a protein/ polymer mixture on a substrate. The nanocomposites were prepared by the addition of gold nanoparticles containing protein molecules, which could easily diffuse into the hydrogel matrix through pores and spaces.…”

“…Chart listed the general backbone chemical structure of the polymers that were mostly used for the synthesis of gel-nanocomposites. Tanaka et al assembled three-dimensional nanostructured protein-based hydrogel composed of “loosely” interconnected protein–polymer hybrid nanoparticles using different proteins (streptavidin, protein A, and Mouse IgG) and a polymer poly­[( N -acryloylmorpholine)- co -( N -acryloxysuccinimide)] . The protein hydrogel has been accomplished by spotting a protein/polymer mixture on a substrate.…”

N-Hexyl-4-aminobutyl glycosides for investigating structures and biological functions of carbohydrates

“…29 The binding between 9 and an Erythrina cristagalli lectin (ECA) was observed using a three-dimensional nanostructured protein hydrogel (3D NPH). 30 We performed the assay using the 3D NPH anticipating the future need for small-scale binding assays. The ECA-containing 3D-protein matrix, prepared by mixing ECA and a copolymer of N-acrylmorpholine and N-acyloxysuccinimide in a phosphate buffer, was found to enable the measurement of lactoside-ECA binding events on amino-functionalized glass using a few picomoles of 9.…”

“…Assays were performed on a micro-well slide (1.0-mmf type1 APS(aminopropylsilyl)-coated glass; Matsunami Glass Ind., Ltd.). To a solution of ECA (10 mL; the amount of ECA was varied in the range 1.76 ¥ 10 -4 -5.50 ¥ 10 -6 M) was added a solution of poly[(N-acrylmorpholine)-co-(N-acyloxysuccinimide)] 30 (1.0 mL, 1% in H 2 O). The mixture was deposited on each well and was kept under wet conditions for 2 h at 37 • C. The 3D-NPH areas were rinsed with 1 M ethanolamine and H 2 O (three times).…”