| Summary: | 碩士 === 國立中正大學 === 化學暨生物化學研究所 === 101 === Knee osteoarthritis (Osteoarthritis, OA), common to people over fifty years old, is the most common form of arthritis in the elderly. Many factors can lead to the degeneration of the knee cartilage, including overuse, injury and hereditary. Joints suffering long periods of gravity and overuse often lead to the cartilage degradation and joint space narrowing.
MMP-13 (Matrix metalloproteinase-13, MMP-13), with a molecular weight of about 52 kDa, is one member of the series of MMPs (matrix metalloproteinases, MMPs). Past studies have shown that the MMP-1, MMP-3 and MMP-13 concentrations on the cartilage in OA patients, are higher than normal.
In this study, we modified the aqueous phase gold nanoparticles on optical fiber probes for analysis of real biological samples by the fiber-optic particle plasmon resonance (FO-PPR) sensor.
The FO-PPR sensor has many advantages such as label-free, high sensitivity, real-time, and low-cost. It offers a potential to monitor biomarkers of various diseases. In this work, we focuses on osteoarthritis concerned biomarker, MMP-13.
First, we tested the sensor with different concentrations of sucrose solutions to determine the sensor resolution and sensor sensitivity. Experimental results show that the sensor resolution can reach 3.78 × 10-6 RIU and the senor sensitivity can reach 4.87 AU / RIU. For conjugation of a biorecognition molecule on the gold nanoparticle surface, we modified a mixed self-assembled monolayer (Mixed SAM) on the surface of the gold nanoparticles, then further conjugate OVA or anti-MMP-13 on the optical fiber probe. For anti-OVA standards, a limit of detection (LOD) of 9.36× 10-7 g/ml was obtained. For MMP-13 standards, the sensor response versus log concentration of MMP-13 is linear (R = 0.9978) and a LOD of 1.35× 10-10g / ml was obtained, which is similar to that by ELISA (Enzyme-linked immunosorbent assay, ELISA).
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