Enhancement of sensitivity of SPR protein microarray using a novel 3D protein immobilization

• Published in: Colloids and surfaces, B, Biointerfaces Vol. 70; no. 2; pp. 259 - 265
• Main Authors: Tanaka, Hiroyuki, Hanasaki, Minako, Isojima, Tatsushi, Takeuchi, Hisao, Shiroya, Toshifumi, Kawaguchi, Haruma
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2009
• Subjects: Animals; Biosensing Techniques - methods; Cross-Linking Reagents - chemistry; Cross-Linking Reagents - pharmacology; Hydrogel; Hydrogel, Polyethylene Glycol Dimethacrylate - chemistry; Hydrogels - chemistry; Imaging, Three-Dimensional - methods; Immobilized Proteins - chemistry; Ligands; Mice; Microscopy, Electron, Transmission - methods; Polymers - chemistry; Protein Array Analysis - methods; Protein immobilization; Protein microarray; Reactive polymer; SPR biosensor; Streptavidin - chemistry; Surface Plasmon Resonance; Protein immobilization; Protein microarray; Hydrogel; Reactive polymer; SPR biosensor
• ISSN: 0927-7765; 1873-4367
• DOI: 10.1016/j.colsurfb.2008.12.037

We discovered a novel method to prepare a protein-based hydrogel, that is, a “Three-Dimensional Nanostructured Protein Hydrogel (3D NPH)”, which is composed of protein–polymer hybrid nanoparticles. In this study, we propose a novel protein microarray whose 3D NPH spots were prepared by dispensing a small volume of the solution of protein–polymer mixture on a substrate. The dispensed solution had a short time for cross-linking before its drying-up and the resulting 3D NPH had loosely cross-linked, thin spongy structure. Therefore, the reaction ratio between ligands and analytes was drastically improved in this system compared with the large volume system for Surface Plasmon Resonance (SPR) protein microarray.