A mixed alkanethiol based immunosensor for surface plasmon field-enhanced fluorescence spectroscopy in serum

Abstract

This paper describes a simple and sensitive immuno-based biosensor for interference-reduced detection of C-reactive protein (CRP) in serum. The detection was performed by using a non-competitive sandwich immunoassay in combination with surface plasmon field-enhanced fluorescence spectroscopy (SPFS). CRP is an important marker for the diagnosis of inflammatory processes and cardiovascular diseases (CVD). It is nowadays detected by high-sensitivity enzyme-linked immunosorbent assays (ELISA) in blood serum. CRP was used as a model analyte in this work because it is well-characterized. However, interfering effects of matrix components affect the limit of detection (LOD) and quantification (LOQ) in general. Therefore, the availability of fast, sensitive and robust analytical methods is of major interest. A number of biosensor approaches have been described already, but only a few have demonstrated their usefulness in authentic samples such as serum. Thus our aim was to develop a simple and sensitive immunoassay-based biosensor for an interference-reduced detection of CRP in serum with surface plasmon enhanced fluorescence spectroscopy (SPFS). LODs and LOQs were experimentally determined both for CRP spiked buffer and serum. SPFS in combination with our biosensor allows sensitive analysis of CRP, achieving in buffer a LOD of 0.016 μg mL⁻¹ and a LOQ of 0.049 μg mL⁻¹. In serum the accomplished LOD was 0.026 μg mL⁻¹ and the LOQ was found to be 0.08 μg mL⁻¹. These low LODs and LOQs demonstrate the applicability of the designed biosensor for qualitative and semi-quantitative analysis of trace amounts of substances in very small sample volumes of body fluids.