Synthesis and Optimization of Fab Antibody Specific to Aflatoxin B1 Conjugated to Gold Nanoparticles
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Abstract
Introduction: Gold nanoparticles was one of the perfect metals for studies the optical properties and most widely applied in biomedicines, biotechnology and diagnostic applications. Fab fragments of antibody specific to aflatoxin B1 were preformed in our laboratory. The Fab-AFB1 coated gold is affected by multiple factors. The optimal conditions were studied for preparation the Fab-AFB1 conjugated gold nanoparticles. The conjugated particles could be used to detect aflatoxin B1 on the basic of the optical property of gold nanoparticles. Materials and methods: in this study, we synthesized the colloidal gold solution by Turkevich method. We studied the effect of pH and concentration of Fab-AFB1 antibody on conjugated in the gold nanoparticles. After inducing aggregation with salt, color changes and UV-absorbance were used to determined for the optimal conditions. Indirect-ELISA was used to measured the unconjugated antibody in conjugation solution for confirming conjugation reaction. Results: The maximum absorption spectra (Xmax, A520>) of coated gold nanoparticle increases from 520 to 531 nm. Increasing pH does not affect on size and the color shown. The optimum concentration was conjugation in molar ratio 0.71. Less conjugated antibody could not prevent salt-induced aggregation. Indirect-ELISA reveal the saturation concentration points above 3.53 in molar ratio. Conclusion: Gold nanoparticles conjugated with Fab-AFB1 antibody specific to aflatoxin B1 is successfully prepared and could be potentially used for versatile application.
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