Novel Biosensor–Based Analytic Device for the Detection of Anti–Double-Stranded DNA Antibodies

Abstract

Background:Patients with systemic lupus erythematosus (SLE) develop a wide variety of serologic manifestations, including double-stranded DNA autoantibodies (anti-dsDNA). The determination of the potentially pathogenic autoantibodies is diagnostically relevant. Methods: We developed a novel surface plasmon resonance (SPR) biosensor chip for studies of dsDNA and anti-dsDNA binding. A synthetic oligonucleotide was coupled to biotinylated human transferrin, hybridized with the complementary antistrand, and ligated with a human recombinant dsDNA fragment 233 bp in length. After surface immobilization of this antigenic construct, diluted sera from SLE patients and healthy donors were analyzed with the resulting SPR biosensor system. Results: This SPR biosensor allowed specific detection of anti-dsDNA. In pilot experiments, sera from SLE patients were distinguished from control sera. We also confirmed the specificity of this biosensor by supplementing anti-dsDNA-positive sera with salmon sperm DNA, which blocked the surface binding of antidsDNA in a concentration-dependent manner. Conclusions: An SPR biosensor monitors interactions in real time under homogeneous conditions, providing information about binding kinetics and affinities. Its applicability critically depends on the design of the

Autoantibodies to double-stranded DNA-Intermethod comparison between four commercial immunoassays and a research biosensor-based device

“…Also, our SPR biosensor marks a new approach with promising possibilities. 10,14 The Farr assay is still considered as the gold standard for clinicians. Advantageous is the high specificity for diagnosing SLE and the discrimination from other autoimmune diseases.…”

Comparison and evaluation of different methodologies and tests for detection of anti-dsDNA antibodies on 889 Slovenian patients’ and blood donors’ sera

“…There were also reports on a novel surface plasmon resonance biosensor chip analytic method (29) and an innovative quantitative electrochemical detection of anti-DNA antibodies (30). Just recently, a novel method using electrophoretic mobility shift assay was reported by Keyhani et al (31), which showed in vivo and in vitro detections of a complex formation between plasma membrane DNA and IgG from SLE patients.…”

Silicon and Silicon-Related Surfaces for Biosensor Applications

“…Surface plasmon resonance (SPR) has been successful so far for many reasons include the easy cleaning and the infinite reuse of the sensor. Thiol based monolayers are normally grafted to gold or silver surfaces allow specific detection of DNA [Buhl, et al, 2007] or organic pollutants [Farré, et al, 2007& Mauriz, et al, 2006 and pathogens [Chah & Zare, 2008] with coupling of DNA and antibodies. A significant improvement has been achieved with the formation of more multifaceted sensing architectures using the assembly of nanoparticles onto the gold surface of the SPR: the signal amplification caused by the coupling of the SPR signal with the localized surface plasmons of the metal or semiconductor nanoparticles (Fig.…”