Complementation of ELISA and an Interdigitated Electrode Surface in Gold Nanoparticle Functionalization for Effective Detection of Human Blood Clotting Defects

Nanoscale Research Letters

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[1] 0000 0000 9363 8679, grid.430704.4, Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, 01000, Kangar, Perlis, Malaysia;0000 0000 9363 8679, grid.430704.4, Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, 01000, Kangar, Perlis, Malaysia;0000 0000 9363 8679, grid.430704.4, School of Bioprocess Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis, Malaysia;0000 0001 2364 8385, grid.202119.9, Department of Biological Engineering, College of Engineering, Inha University, 402-751, Incheon, Republic of Korea;grid.414902.a, Department of Vascular Surgery, The First Affiliated Hospital of Kunming Medical University, 650032, Kumming, Yunnan, China;grid.414918.1, Department of General Surgery, The First People’s Hospital of Yunnan Province, the Affiliated Hospital of Kunming University of Science and Technology, 650032, Kunming, Yunnan, China;

关键词: ELISA;    Gold nanoparticle;    Factor IX;    Silanization;    Blood clotting;    Human serum;    Interdigitated electrode;   

DOI  :  10.1186/s11671-019-3058-z

来源: publisher

【 摘 要 】

Developing an enhanced diagnosis using biosensors is important for the treatment of patients before disease complications arise. Improving biosensors would enable the detection of various low-abundance disease biomarkers. Efficient immobilization of probes/receptors on the sensing surface is one of the efficient ways to enhance detection. Herein, we introduced the pre-alkaline sensing surface with amine functionalization for capturing gold nanoparticle (GNP) conjugated to human blood clotting factor IX (FIX), and we demonstrated the excellent performance of the strategy. We have chosen the enzyme-linked immunosorbent assay (ELISA) and the interdigitated electrode (IDE), which are widely used, to demonstrate our method. The optimal amount for silanization has been found to be 2.5%, and 15-nm-sized GNPs are ideal and characterized. The limit of FIX detection was attained with ELISA at 100 pM with the premixed GNPs and FIX, which shows 60-fold improvement in sensitivity without biofouling, as compared to the conventional ELISA. Further, FIX was detected with higher specificity in human serum at a 1:1280 dilution, which is equivalent to 120 pM FIX. These results were complemented by the analysis on IDE, where improved detection at 25 pM was achieved, and FIX was detected in human serum at the dilution of 1:640. These optimized surfaces are useful for improving the detection of different diseases on varied sensing surfaces.

【 授权许可】

CC BY