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Optimization Studies on the Manufacturing Process of Colloidal Gold-based Detection for Ractopamine

Author: HuangXiaoMing
Tutor: TangYong
School: Jinan University
Course: Biological Engineering
Keywords: Ractopamine (RAC) Colloidal gold Coupling Nitrocellulose membrane Aging
CLC: TQ421
Type: Master's thesis
Year: 2011
Downloads: 305
Quote: 0
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Abstract


Objective: to the initial establishment of ractopamine colloidal gold strip production process , optimizing the various aspects of the production process of the colloidal gold , including gold solution preparation, preparation of the conjugate of the gold standard , the nitrocellulose membrane processing and storage conditions settings, etc. establish industrialization ractopamine colloidal gold strip production process. Methods: A single - factor method , Preparation of monodisperse colloidal gold process optimization application trisodium citrate reduction by changing the reaction time and the tri-sodium citrate concentration and visible spectrophotometer (400-700nm) and scanning electron microscope observe the uniformity and dispersion of gold particles . Optimization gold nanoparticles RAC antibody conjugated to RAC antibody dilution factor and the coupling time . Drying treatment at 37 ° C and 55 ℃ nitrocellulose membrane was observed within 5 days of the NC membrane RAC antigen and goat anti -mouse secondary antibody fixed case . Colloidal gold strip at 37 ℃ accelerated aging, and saved as a control group, Group 3 ( untreated, heat treatment , freezing process) at room temperature . Optimal conditions in the above the repeatability experiment three times , comparing the differences of different batches of colloidal gold strip . Results: When the trisodium citrate concentration is 5% , the reaction time was 20min , concentrated particle size can be obtained a single dispersion of a good colloidal gold solution . Gold nanoparticles with RAC antibody conjugated coupling time 30min antibody diluted 15 times to achieve the best coupling effect . Nitrocellulose membrane was dried for 4-5 days in the 37 ° C case need to reach a steady state , to 55 ℃ simply time can be dried for 3 days . Aging experimental results show whether processing at room temperature , and within six months the test strip is very stable and did not change significantly , while the effect of the test strip within the next 3 months at 37 ℃ , when the time is extended to 6 months, the test strip the slight decrease in the quality , sensitivity down to 10ng/ml . In the results of the repeated experiments of three times that different batches of the assembly of colloidal gold strip results are different , but the difference was not significant . Conclusion: This study perfect ractopamine colloidal gold strip production process , including the preparation of the gold solution , the preparation of the complex of the gold standard , the NC membrane treatment , and ultimately aging experiments show that the test strip shelf life of at least 1 year , but different batches of colloidal gold strip difference is not significant , with good reproducibility and stability .

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