title: Analysis of C-reactive protein on amide-linked N-hydroxysuccinimide-dextran arrays with a spectral surface plasmon resonance biosensor for serodiagnosis.
abstract: A new label-free array system using amide-linked (AL) NHS-dextran and a spectral SPR biosensor are presented for the high-throughput analysis of C-reactive protein (CRP) in human sera. The AL NHS-dextran layer on the surface of gold arrays was composed of an amide linkage between NHS-modified carboxymethyl-dextran and amine-modified 11-mercaptoundecanoic acid. The topology of the AL NHS-dextran layer was analyzed by atomic force microscopy, and it was found to be superior to the previously used epoxide-linked carboxymethyl-dextran layer in its immobilization of proteins. Specific immunoreactions and a dose-dependent increase of SPR signals were demonstrated on the AL NHS-dextran layer. Then, the label-free array system was successfully applied to the rapid analysis of CRP in 120 human sera. CRP levels in human sera determined by the array-based spectral SPR biosensor showed a good correlation with those determined by the latex-enhanced turbidimetry immunoassay (n = 120, r = 0.945, p < 0.0001). Thus, the array-based spectral SPR biosensor based on the AL NHS-dextran surface is a potential system for rapid and label-free serodiagnosis of human diseases.
authors.affiliation: Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 200-701, South Korea.
authors.forename: Se-Hui
authors.initials: SH
authors.lastname: Jung
authors.forename: Jae-Wan
authors.initials: JW
authors.lastname: Jung
authors.forename: In-Bum
authors.initials: IB
authors.lastname: Suh
authors.forename: Jong Seol
authors.initials: JS
authors.lastname: Yuk
authors.forename: Woo-Jin
authors.initials: WJ
authors.lastname: Kim
authors.forename: Eui Yul
authors.initials: EY
authors.lastname: Choi
authors.forename: Young-Myeong
authors.initials: YM
authors.lastname: Kim
authors.forename: Kwon-Soo
authors.initials: KS
authors.lastname: Ha
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002097:C-Reactive Protein / Q000032:analysis*; D003911:Dextrans / Q000737:chemistry*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D013388:Succinimides / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D003911:Dextrans; D013388:Succinimides; D002097:C-Reactive Protein; C001426:N-hydroxysuccinimide