研究目的
To develop a photoluminescence immunosensor based on bovine leukemia virus proteins immobilized on ZnO nanorods for the determination of specific antibodies against BLV proteins in cattle serum.
研究成果
The developed photoluminescence immunosensor based on gp51&BSA/PAH/ZnO-NR/glass can differentiate between 'BLV-positive' and 'BLV-negative' serums in a dilution range from 1:1000 to 1:100000, with the signal generation primarily based on electrostatic interactions. It offers a rapid, sensitive, and cost-effective method for BLV detection.
研究不足
The mechanism of interaction between biomolecules and ZnO surface is not fully understood. Random orientation of antigens may reduce sensitivity. The system may have limitations in specificity and requires further optimization for broader applications.
1：Experimental Design and Method Selection:
The study involved designing an immunosensor using ZnO nanorods as a transducer, with photoluminescence spectroscopy to detect changes upon antigen-antibody binding. Theoretical models for electrostatic interactions and photoluminescence mechanisms were considered.
2：Sample Selection and Data Sources:
BLV proteins gp51, 'BLV-positive' and 'BLV-negative' cattle serums were purchased from "Leykonad", Poltava, Ukraine. Glass slides, PAH, butanol, and BSA were obtained from Sigma-Aldrich.
3：List of Experimental Equipment and Materials:
Equipment included Hitachi S4800 SEM, LGI-21 nitrogen laser, HR2000+ spectrometer, ALFA300R Raman spectrometer. Materials included ZnO nanorods, PAH, BSA, glass substrates, butanol, NaCl solutions.
4：Experimental Procedures and Operational Workflow:
ZnO nanorods were synthesized and deposited on glass, annealed, and characterized by SEM and Raman spectroscopy. Functionalization involved immobilizing gp51 and BSA on ZnO-NR/glass with or without PAH modification. Incubation with serum samples was done for 20 minutes, followed by washing and PL measurement.
5：Data Analysis Methods:
PL spectra were recorded and normalized. The immunosensor response was calculated using S = 1 - Inorm(C), where Inorm(C) is the normalized PL intensity. Data were averaged from at least three measurements.
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Scanning Electron Microscope
S4800
Hitachi
Imaging of ZnO nanorods on glass substrates
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Optical Fiber Spectrometer
HR2000+
Ocean Optics
Recording photoluminescence spectra
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Nitrogen Laser
LGI-21
Polyaron
Excitation source for photoluminescence spectroscopy
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Raman Spectrometer
ALFA300R
Witek
Raman spectroscopy measurements
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Milli Q-plus System
Milli Q-plus
Millipore
Water purification for solution preparation
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