Abstract
This paper presents a generally applicable approach for the highly specific detection of blood proteins. Thrombin and thrombin-binding aptamers are chosen for demonstration purposes. The sensor was prepared by immobilizing amine-terminated aptamers onto a gold modified surface using a two-step self-assembled monolayer (SAM) immobilization technique and the physical detection is performed using Surface Plasmon Resonance (SPR). The developed sensor has an optimal detectable range of 5–1000 nM and the results show the sensor has good reversibility, sensitivity and selectivity. Furthermore, the sensor shows the potential of being improved and standardized for direct detection of other blood proteins for clinical applications.
Highlights
The direct detection of blood proteins can benefit a number of scientific and clinical applications, such as in monitoring the ratio of specific protein glycation in diabetes [1], biomarkers for drug research and environmental monitoring [2], cancer diagnostics and treatment [3], etc
We report on a 3-mercaptopropionic acid (MPA) self-assembled monolayer (SAM) based coupling approach for a more stable and repeatable modification of the sensor surface
Aptamer solutions were prepared with 1M pH 8 phosphate buffer and 3-mercaptopropionic acid (MPA) solution was prepared in ethanol
Summary
The direct detection of blood proteins can benefit a number of scientific and clinical applications, such as in monitoring the ratio of specific protein glycation in diabetes [1], biomarkers for drug research and environmental monitoring [2], cancer diagnostics and treatment [3], etc. An approach is presented for the detection of specific blood proteins based on custom SPR aptamer modified sensors that can have broad applications. Aptamers are artificial oligonucleotides which can serve as antibody mimics because of their high affinity and selectivity for various target compounds ranging from small molecules, such as drugs and dyes, to complex biological molecules such as enzymes, peptides, and proteins. Some groups have successfully developed aptamer-based biosensors for the detection of proteins [17,18,19,20,21,22,23,24] and related SPR based sensors have been reported [25,26].
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