基于多肽的真菌毒素全抗原模拟物结构重建及其免疫学分析特性的分子机制

全抗原(包被及竞争抗原)的化学合成制备已成为建立真菌毒素免疫学分析体系及规模化应用的技术瓶颈之一。本研究拟在获得真菌毒素半抗原模拟表位的前期工作基础上，以多肽分子为核心元件，开展真菌毒素全抗原模拟物的结构重建。通过半抗原突变库的构建及差异化淘选，获取与抗体具有不同结合平衡常数Ka数值的多肽分子；并用分子电距矢量（MEDV）表征多肽的分子结构特征并构建其定量构效关系模型，分析其空间结构域变化对其亲和能力的影响；同时采用分子模拟方法将多肽分子与不同的骨架蛋白进行模拟嵌合，构建基于肽模拟的真菌毒素全抗原模拟物；探析模拟物的基序、柔性度以及空间结构域对抗体分子识别取向、特异性及灵敏度的影响机制；在此基础上，通过基因工程的方法进行真菌毒素全抗原模拟物的生物合成，探析其免疫分析性能的稳定性，并应用于不同免疫学检测模式。研究成果对其它有毒小分子物质全抗原模拟物的生物制备也具有积极的示范意义。

Chemical synthesis of mycotoxin conjugates is the technique bottleneck of the development of immunoassay method and its large-scale application in detection of mycotoxins. In this study, we intend to use peptide as core element to develop and reconstruct of mycotoxin conjugates mimics based on our previous research works of mycotoxin mimotope. First, we are trying to construct a mycotoxin mimotope mutant library and select peptide molecules with different affinity constant of anti-mycotoxin antibodies by various biopanning strategy. After that, molecular electronegativity -distance vector(MEDV) is proposed to characterize the structure of peptide molecules and elucidate the structue-activity relationship .In addition, peptide molecules and scaffold were combined by computer molecular modeling to construct mycotoxin conjugate peptide mimics, and the influence mechanism of motif , flexibility and conformation domain on antibody detection performance were analyzed. Furthermore, to large-scale preparation of peptide mimics, the gene of coding peptide will be transferred to express vector and express peptide-scaffold fusion protein and applied in different type immunoassay . The final goal of this study is to provide the theoretical and methodological support for the peptide applied to biosynthesis of complete conjugates of toxic small molecules.