title: Establishment of N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC) modified biochip enabling concurrent detection of serum infectious antibodies in neuroborreliosis.
abstract: In this study, we developed a novel protein biochip that was modified with N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC) and specialized for concurrent detection of serum IgG and IgM antibodies against Borrelia burgdorferi antigens, flagellin, outer surface protein C (OspC) and variable major protein-like sequence (VlsE) in the patients with neuroborreliosis (NB), respectively. Surface chemical characteristics of the biochips were validated with atomic force microscope (AFM) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). The visualized detection limit for IgG antibodies against flagellin, OspC and VlsE antigens on the biochip were 0.78 µg/ml, 0.78 µg/ml and 1.56 µg/ml, respectively. Finally, serum IgG and IgM antibodies in 72 patients with NB and 188 healthy individuals were tested on the biochip. The seroimmunological outcome by the biochip were evaluated in comparison with enzyme linked immunosorbent assay (ELISA) assay. The results demonstrated that the prevalences of IgG and IgM antibodies in the cases were 41.7%, 63.9% to flagellin; 20.8% and 51.4% to OspC and 76.4%, 62.5% to VlsE, respectively. Utilization of the biochip in detection IgM antibody against flagellin was compatible with ELISA assay (R(2)=0.849). Thus, the protein biochip would provide a potential platform not only for enabling detection of corresponding antibodies directed against B. burgdorferi antigens, but also for monitoring course of the disease.
authors.affiliation: Department of Biology, School of Life Sciences, Anhui Medical University, Meishan 81, Hefei 230032, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Ye
authors.affiliation: Department of Biology, School of Life Sciences, Anhui Medical University, Meishan 81, Hefei 230032, PR China.
authors.forename: Na-Li
authors.initials: NL
authors.lastname: Huang
authors.affiliation: MVZ, ACURA Kliniken, Baden-Baden 76530, Germany.
authors.forename: Xue-Ling
authors.initials: XL
authors.lastname: Ma
authors.affiliation: Sektion Experimentelle Anästhesiologie, Universitätsklinikum Ulm, Albert Einstein Allee 23, Ulm 89081, Germany. Electronic address: marion.schneider@uniklinik-ulm.de.
authors.forename: Marion
authors.initials: M
authors.lastname: Schneider
authors.affiliation: Research Centre for Biomimetic Functional Materials and Sensing Devices, Institute of Intelligent Machines, Chinese Academy of Science, Hefei 230031, PR China.
authors.forename: Xin-Jiu
authors.initials: XJ
authors.lastname: Huang
authors.affiliation: Sektion Experimentelle Anästhesiologie, Universitätsklinikum Ulm, Albert Einstein Allee 23, Ulm 89081, Germany; Department of Pathology, Anhui Medical University, Hefei 230032, PR China. Electronic address: weidongdu@hotmail.com.
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Du
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification; D015374:Biosensing Techniques; D005408:Flagellin / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D008074:Lipoproteins / Q000276:immunology / Q000302:isolation & purification; D020852:Lyme Neuroborreliosis / Q000097:blood*; D008301:Maleimides / Q000737:chemistry*; D040081:Protein Array Analysis
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008074:Lipoproteins; D008301:Maleimides; C079616:OspC protein; C105546:VlsE protein, Borrelia burgdorferi; D005408:Flagellin; C025285:N-(4-carboxycyclohexylmethyl)maleimide N-hydroxysuccinimide ester
keywords: ELISA; Flagellin; N-Succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC); Neuroborreliosis; Outer surface protein C (OspC); Protein biochip; Variable major protein-like sequence (VlsE)