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首页> 外文会议>International Conference on Bioscience, Biotechnology, and Biometrics >Purification of polyclonal antibody against pork extracts antigens using Protein A Column as material for developing halal food detection kit
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Purification of polyclonal antibody against pork extracts antigens using Protein A Column as material for developing halal food detection kit

机译：用蛋白质a柱作为开发清真食品检测试剂盒的材料纯化猪肉抗猪肉提取抗原的纯化

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The main objective of this study was to purify polyclonal antibodies against pork extract as raw material for developing diagnostic kits for halal food made from animal products. Antibodies were obtained from rabbit serum which was immunized with raw pork extract, and which had been heated at 70°C, 80°C and 100°C. The purification process was carried out in two stages namely using 33% ammonium sulfate and continued with Protein A column affinity. The antibody purification results were analyzed using SDS-PAGE, while the antibody activity was tested using the western blot method on raw meat extracts and on those heated at 70°C , 80°C, and 100°C. The results of this study showed that antibodies purified with a combination of ammonium sulfate and Protein A column affinity produced relative purity of antibodies up to 97.6 per cent compared to those purified using ammonium sulfate alone. Western blot analysis showed that the purified antibodies were able to detect raw as well as heated pork extracts up to 70°C at a total protein content of 20 micro grams of meat extract. Antibodies obtained from immunizations using meat extracts that have been heated at 70°C gave the same potential as antibodies from rabbits immunized using raw meat extract, i.e. can only detect antigens of raw meat and antigens of pork heated at 70°C. From the results of this study it can be concluded that purification of antibodies using a combination of ammonium sulfate and Protein A column is effective in producing polyclonal antibodies with relatively high purity levels. In the future, further research needs to be carried out on the development of immunodiagnostics using polyclonal antibodies purified from animals immunized with pork extract that has been heated at a temperature of 70°C or more.
机译：本研究的主要目的是将多克隆抗体净化猪肉提取物作为制造由动物产品制成的清真食品的诊断试剂盒。用兔血清获得抗体，其用原料猪肉提取物免疫，并在70℃，80℃和100℃下加热。纯化过程在两个阶段中进行，即使用33％硫酸铵并继续蛋白质柱亲和力。使用SDS-PAGE分析抗体纯化结果，同时使用Western Blot方法在原肉提取物上进行抗体活性，并在70℃，80℃和100℃下加热的那些。该研究的结果表明，与使用单独纯化的纯化的那些，用硫酸铵和蛋白质的组合纯化的抗体和蛋白质柱亲和力产生的相对纯度高达97.6％。 Western印迹分析表明，纯化的抗体能够以20微克肉提取物的总蛋白质含量检测到最良好的猪肉和高达70℃的加热猪肉提取物。使用在70℃下被加热的肉提取物获得的免疫接种获得的抗体作为使用生肉提取物免疫的兔子的抗体，即只能检测在70℃下加热的猪肉的抗原和猪肉的抗原。从该研究的结果，可以得出结论，使用硫酸铵和蛋白质的组合纯化抗体，柱是有效地产生具有相对高纯度水平的多克隆抗体。在未来，需要进一步研究使用从用猪肉提取物免疫的动物纯化的多克隆抗体在猪肉提取物中被加热在70℃或更高的温度下进行免疫抗体的发展。

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