Biological activity of chemically immobilized enzymes on nanostructured solid supports is determined by chemical structures of functional groups on the solid surfaces, which may affect the conformation of the proteins at varied levels. Thus, control of the protein conformation on the surfaces and interaction between proteins and the solid surfaces play a crucial role in fabrication of biosensors by using immobilized enzymes. Here, we fabricated varied nanofibrous membranes from these polymers, including silica, poly(vinyl alcohol-co-ethylene) (PVA-co-PE), and polystyrene (PS), which have different hydrophilicity and protein compatibility. Then, selected antibodies (Abs) were immobilized onto these nanostructured polymers via varied polyethylene glycol (PEG) chain lengths and functional groups. Disuccinimidyl carbonate (DSC) demonstrated high interference to conformation of antibodies because of its high reactivity to amine groups on N-terminal of proteins. Cyanuric acid (CC) was found to provide the most stable covalent bonds with the protein and light influence on protein conformation. Fluorescence spectroscopy, circular dichroism (CD) spectroscopy, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and bio-activity to specific antigens were used in determination of bioactivities of the proteins immobilized on the nanofibrous membranes.

Study on protein conformation and bio-activity of antibodies immobilized on varied nanofibrous membranes

253rd National Meeting (2017)

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