Sintesis antigen AFB1-BSA dan konjugasi antibodi anti AFB1-BSA dengan nanopartikel emas sebagai pereaksi imunostrip
Abstract
Aflatoksin B1 (AFB1) often contaminates a great variety of foods and animal feeds that will be dangerous if con-sumed by humans or animals. Rapid detection techniques that can be used in the field is really needed to monitor AFB1 contamination. The aims of this study were to perform synthesis of AFB1-BSA antigen and conjugation of antibody against AFB1-BSA to gold nanoparticle as immunostrip-test reagents. The AFB1-CMO was identified on TLC and AFB1-BSA was characterized using SDS PAGE. The AFB1-CMO formation indicated as a blue spot at 0.45 retention factor (Rf) on TLC and the AFB1-BSA antigen revealed as a single band protein at about 72 kDa molecular weight on the SDS PAGE. Conjugation of antibody against AFB1-BSA to gold nanoparticle resulted in the formation of red-dish purple compound which can be used for the detection of AFB1 on immunostrip. The optimum composition achieved in concentration of AFB1-BSA 1-1.5 mg/ml, IgG anti rabbit 0.1 mg/ml, and antibody against AFB1-BSA-gold nanoparticle conjugate in 0.5x0.5 cm² area characterized by the establishment of two reddish purple lines in the test and control zone.
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References
AOAC. 1995. Official Methods of Analysis of AOAC International. Maryland: AOAC International.
Chiao DJ, Shyu RH, Hu Cs, Chiang HY, Tang SS. 2004. Colloidal gold based immunochromatographic assay for detection of botulinum neurotoxin type B. Journal of Chromatography B. 809(1): 37-41.
Chu FS, Hsia SMT, Sun PS. 1977. Preparation and characterization of AFB1-oxime. Association Official Analytical Chemistry. 60:791-794.
Hames BD. 1998. Gel Electrophoresis of Protein 3rd ed. New York. Oxford University Press.
[IARC] International Agency of Research on Cancer. 2002. Some traditional herbal medicines, some mycotoxins, naphthalene and styrene. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. 82: 1-556.
Laemmli UK. 1970. Cleavage of Structural Protein during the assembly of the head of bacteriophage T4. Nature. 227(5259): 680-685.
Li P, Zhang Q, Zhang W. 2009. Immunoassays for aflatoxins. Trends in Analitical Chemistry. 28(9): 1115-1126.
Liu BH, Yu-Tien Hsu, Chuan-Chen Lu, Feng-Yih Yu. 2013. Detecting AFB1 in foods and feeds by using sensitive rapid enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip. Food Control. 30(1):184-189.
Maciorowskia KG, Herreraa P, Jonesb FT, Pillaia SD, Rickea SC. 2007. Effects on poultry and livestock of feed contamination with bacteria and fungi. Animal Feed Science and Technology. 133 (1-2): 109–136.
Saini SS, Kaur A. 2012. Aflatoxin B1: Toxicity, characteristics and analysis: Mini review. Global Advanced Research Journal of Chemistry and Material Science. 1(4): 63-70.
Zheng MZ, Richard JL, Binder J. 2006. A review of rapid methods for the analysis of mycotoxins. Mycopatologia 161(5): 261-273.
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