Abstract
Two label-free biosensor platforms, Resonance Waveguide Grating (RWG) and Surface Plasmon Resonance (SPR), were used to rank a large panel of anti-dengue virus NS1 antibodies. Dengue non-structural 1 (NS1) protein is an established serological marker for the early detection of dengue infection. A variety of commercial dengue NS1 antigen capture immunoassays are available in both ELISA and lateral flow format. However, there is a significant scope to improve both the sensitivity and the specificity of those tests. The interactions of antibody (Ab)-antigen (Ag) were profiled, with weak interactions (KD= 1–0.1 μM) able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR. There were significant differences in the absolute affinities determined by the two technologies, and there was a poor correlation between antibodies best ranked by RWG and the lower limit of detection (LLOD) found by ELISA. Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.
Highlights
Dengue is a major public health concern worldwide
Polyclonal anti-dengue non-structural 1 (NS1) antibodies used in the ELISA lower limit of detection (LLOD) study were provided by Alere, Australia
NS1 mAbs that possessed immuno-reactivity in any of these three immunoassays were included in the Resonance Waveguide Grating (RWG) and Surface Plasmon Resonance (SPR) assays
Summary
Dengue is a major public health concern worldwide. It is estimated that more than 2.5 billion people live in endemic areas, with 50 to 100 million people suffering from dengue infection, resulting in approximately 25,000 deaths reported annually [1]. Despite efforts focused on vector control, the incidence of dengue has increased markedly over the past 50 years [2]. As a licensed vaccine or anti-viral therapeutics for dengue remain elusive, the control of dengue still relies heavily on an early and accurate diagnosis. The routine diagnostic methods of dengue include a combination of clinical signs, viral isolation [3], reverse transcription polymerase chain reaction (RT-PCR) [4], and serology [5]. These methods can provide accurate diagnosis; none of them are sufficiently sensitive and specific to be used as a stand-alone diagnostic tool [6]
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