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Depletion of algal toxin-contaminated water using selective biofilters based on plant-produced antibodies (plantibodies)

使用基于植物产生的抗体（plantibodies）的选择性生物过滤器去除藻类毒素污染的水

基本信息

批准号：
225858526
负责人：
Professor Dr. Dietmar Knopp
金额：
--
依托单位：
Department für Biotechnologie (DBT)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2013
资助国家：
德国
起止时间：
2012-12-31 至 2016-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/225858526?language=en
关键词：
Depletion algal toxin contaminated water

项目摘要

Although the use of genetically modified plants for bioremediation, or the in situ cleaning of contaminated sites, has been known for quite some time, little attention has so far been paid to the production of antibodies in plants and their ex vivo application in selective depletion. Therefore, highly affine and specific antibodies against algal toxins using microcystin as an example will be produced in plants at low cost within this research project. The basis is a monoclonal antibody (Mab 10E7, species: mouse) generated in a former research project. The sequence of the variable domains will be determined, optimised for plants and subcloned into suitable plant transformation vectors, which already contain constant antibody sequences. In addition, a scFv fragment containing different tag sequences and fusion proteins will be constructed. Leaf-based (tobacco) as well as seed-based (barley) systems will be used.Affinity-purified plant-produced antibodies (plantibodies) will be characterised in detail for their binding properties using microtitre plate-ELISA and surface plasmon resonance (SPR). The monoclonal mouse antibody will be used as reference. To assure cost-efficiency for future applications, roughly purified fractions (sequential pH and temperature treatment followed by filtration) will be tested for the upscaling. Following immobilisation of the plantibody fractions on suitable substrates, for instance membranes, porous polymer monoliths or in porous glasses, their application for depletion will be defined using model water samples spiked fortified with microcystins.

虽然利用转基因植物进行生物修复或污染部位的原位清理已经知道了很长一段时间，但到目前为止，人们对植物中抗体的产生及其在选择性消耗中的体外应用还很少关注。因此，本研究项目将以微囊藻毒素为例，在植物中以低成本生产高度仿合的和特异性的抗藻毒素抗体。其基础是一种在以前的研究项目中产生的单抗(Mab 10E7，物种：小鼠)。可变区的序列将被确定，针对植物进行优化，并亚克隆到合适的植物转化载体中，这些载体已经包含恒定的抗体序列。此外，还将构建包含不同标签序列和融合蛋白的单链抗体片段。将使用基于叶片(烟草)和基于种子(大麦)的系统。亲和力纯化的植物产生的抗体(植物抗体)将使用微滴定板-ELISA和表面等离子共振(SPR)详细表征其结合特性。以小鼠的单抗为参照物。为了确保未来应用的成本效益，粗提级分(顺序的pH和温度处理，然后过滤)将为扩大规模进行测试。将植物体组分固定在合适的基质上后，例如薄膜、多孔聚合物整体或多孔玻璃，将使用添加了微囊藻毒素的模型水样来定义它们在耗尽方面的应用。