Abstract
Design of an optimal surface biofunctionalization still remains an important challenge for the application of biosensors in clinical practice and therapeutic follow-up. Optical biosensors offer real-time monitoring and highly sensitive label-free analysis, along with great potential to be transferred to portable devices. When applied in direct immunoassays, their analytical features depend strongly on the antibody immobilization strategy. A strategy for correct immobilization of antibodies based on the use of ProLinker™ has been evaluated and optimized in terms of sensitivity, selectivity, stability and reproducibility. Special effort has been focused on avoiding antibody manipulation, preventing nonspecific adsorption and obtaining a robust biosurface with regeneration capabilities. ProLinker™-based approach has demonstrated to fulfill those crucial requirements and, in combination with PEG-derivative compounds, has shown encouraging results for direct detection in biological fluids, such as pure urine or diluted serum. Furthermore, we have implemented the ProLinker™ strategy to a novel nanoplasmonic-based biosensor resulting in promising advantages for its application in clinical and biomedical diagnosis.
Highlights
During the last decades, biosensors have emerged as essential tools in biomedical applications, offering exceptional advantages over conventional clinical techniques for diagnostics and therapy monitoring
ProLinkerTM strategies showed higher signal response, being the ProLinkerTM 10 times higher than the covalent method using the same concentration of antibody, which indicates a significant higher antibody binding efficiency
Strong and stable complexes are formed with Arg and Lys, by means of electrostatic interactions. These interactions ensure the formation of strong complexes with aminated proteins such as antibodies
Summary
Biosensors have emerged as essential tools in biomedical applications, offering exceptional advantages over conventional clinical techniques for diagnostics and therapy monitoring. Biosensing platforms and optical biosensors in particular provide rapid, reproducible and highly sensitive detection. They allow label-free analysis (i.e., photonic and plasmonic biosensors) and real-time monitoring of biological events. They have shown exceptional potential for integration in lab-on-a-chip devices [1,2]. The direct immunoassay should be the preferred format for diagnosis since it permits one-step analyte detection and, ideally, direct measurements from biological fluids To fulfill this unmet need and to improve the overall efficiency of the process, many biofunctionalization strategies for antibody immobilization have been studied [4,5].
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