Sintesis antigen AFB1-BSA dan konjugasi antibodi anti AFB1-BSA dengan nanopartikel emas sebagai pereaksi imunostrip

Ita Krissanti
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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.
黄曲霉素B1 (AFB1)经常污染各种各样的食物和动物饲料,如果被人类或动物食用将是危险的。监测AFB1污染确实需要能够在现场使用的快速检测技术。本研究的目的是合成AFB1-BSA抗原,并将抗AFB1-BSA抗体偶联到金纳米颗粒上作为免疫试纸试剂。TLC鉴定AFB1-CMO, SDS - PAGE鉴定AFB1-BSA。在薄层色谱上,AFB1-CMO形成为0.45保留因子(Rf)的蓝色斑点;在SDS PAGE上,AFB1-BSA抗原显示为分子量约72 kDa的单带蛋白。将抗AFB1- bsa抗体与金纳米颗粒偶联形成红紫色化合物,可用于AFB1免疫条检测。最佳组合为AFB1-BSA浓度1 ~ 1.5 mg/ml,抗兔IgG浓度0.1 mg/ml,抗AFB1-BSA-金纳米颗粒偶联抗体浓度0.5x0.5 cm²,在试验区和控制区建立两条红紫色线。
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