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.
Keywords: antibody immobilization; ProLinker™; direct immunoassay; SPR biosensor; nanoplasmonic biosensor; gold nanodisks; clinical diagnosis
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Soler, M.; Estevez, M.-C.; Alvarez, M.; Otte, M.A.; Sepulveda, B.; Lechuga, L.M. Direct Detection of Protein Biomarkers in Human Fluids Using Site-Specific Antibody Immobilization Strategies. Sensors 2014, 14, 2239-2258.
Soler M, Estevez M-C, Alvarez M, Otte MA, Sepulveda B, Lechuga LM. Direct Detection of Protein Biomarkers in Human Fluids Using Site-Specific Antibody Immobilization Strategies. Sensors. 2014; 14(2):2239-2258.
Soler, Maria; Estevez, M.-Carmen; Alvarez, Mar; Otte, Marinus A.; Sepulveda, Borja; Lechuga, Laura M. 2014. "Direct Detection of Protein Biomarkers in Human Fluids Using Site-Specific Antibody Immobilization Strategies." Sensors 14, no. 2: 2239-2258.