聚合物纳米粒子对毒素蛋白的抑制性能研究  

The study on the inhibitory properties of polymer nanoparticles to toxin proteins

作　　者：高胜 李晓晓 鲁希华[1] GAO Sheng;LI Xiao-xiao;LU Xi-hua(College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China)

机构地区：[1]东华大学化学化工与生物工程学院,上海201620

出　　处：《应用化工》2021年第11期2929-2933,2940,共6页Applied Chemical Industry

基　　金：国家自然基金(51473032);国家重点研发项目(2017YFB0309100)。

摘　　要：采用乳液聚合法制备了一系列基于N-异丙基丙烯酰胺(NIPAm)含有不同功能性单体的纳米凝胶粒子(NPs),使用透射电子显微镜和动态光散射仪表征其形貌和分散性能。通过蛋白酶实验和SDS-PAGE电泳实验,筛选对尖吻蝮蛇毒蛋白有抑制作用的功能性纳米粒子。在血清存在的环境中进一步探索了NPs对蛇毒的抑制作用,并采用MTT细胞实验,探究NPs的生物相容性。结果表明,合成的多种NPs分散性良好(PDI<0.1);蛋白酶实验发现,含有N-丙烯酰基苯丙氨酸(Aphe)的poly(NIPAm-Aphe)NPs对尖吻蝮蛇毒的抑制作用较好,可以达到80%;且电泳实验进一步证明了NPs对蛇毒的吸附作用。在血清存在的环境中(<40 mg/mL时),NPs对尖吻蝮蛇毒的抑制作用仍然维持可观水平(50%)。细胞活性实验证明,NPs生物相容性良好,细胞存活率均可达到95%以上,具备作为人工合成抗体的应用潜力。A library of NIPAm-based NPs containing different monomers was synthesized by emulsion copolymerization to screen the NPs that have inhibitory effects on Agkistrodon acutus venom.Morphology and particles size distribution were characterized by transmission electron microscopy(TEM)and dynamic light scattering(DLS).The inhibition effects on venom were characterized by protease and SDS-PAGE experiments.And MTT assays were used to investigate the biocompatibility of NPs.The results showed that synthesized NPs had well distribution(PDI<0.1).And poly(NIPAm-Aphe)NPs containing N-acryloylphenylalanine(Aphe)had relatively excellent inhibition effects on venom,up to 80%.SDS-PAGE experiment further proved the adsorption of snake venom by NPs.Besides,protease experiment showed the nanoparticles inhibition effect on Agkistrodon acutus venom still got to 50%even in the presence of serum(<40 mg/mL).Moreover,cell viability experiments proved that NPs have no obvious cytotoxicity,and cell survival rate can reach more than 95%,the optimized NIPAm-Aphe copolymer nanogels will be promising to be used for synthetic antibody.

关 键 词：纳米颗粒 尖吻蝮蛇 生物大分子 蛋白酶实验 

分 类 号：TQ31[化学工程—高聚物工业] O632[理学—高分子化学]