Pei Shuailong, Sun Wanting, Han Qiang, Wang Haibo, Liang Qionglin
College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian, 116000, PR China; MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Department of Chemistry, Laboratory of Flexible Electronics Technology, Center for Synthetic and Systems Biology, Tsinghua University-Peking University Joint Centre for Life Sciences, Tsinghua University, Beijing, 100084, PR China.
MOE Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Department of Chemistry, Laboratory of Flexible Electronics Technology, Center for Synthetic and Systems Biology, Tsinghua University-Peking University Joint Centre for Life Sciences, Tsinghua University, Beijing, 100084, PR China.
Talanta. 2024 May 15;272:125790. doi: 10.1016/j.talanta.2024.125790. Epub 2024 Feb 17.
The investigation of exosomes holds substantial importance in the field of disease diagnosis and prognosis, where in the rapid and low-loss isolation of exosomes emerges as a crucial step owing to their exceedingly low concentration in bodily fluids. Herein, bifunctional immunoaffinity magnetic nanoparticles (BI-NPs) were successfully constructed by binding Ti and antibody to magnetic nanoparticles through host-guest interaction. Exosomes in the cell culture supernatant could be effectively captured by the BI-NPs and then gently eluted with α-CD and 10% ammonia solution. Following the elution process, the content of captured exosome protein was determined to be up to 97 μg/mL. A total of 2822 protein groups were identified in the exosomes isolated by BI-NPs, 1060 protein groups were derived from exosomes. At the same time, the mass of exosome protein obtained by BI-NPs was more than 2 times that of UC isolation, and the above results indicated that BI-NPs had high purity enrichment performance. This is attributed to the fact that BI-NPs combine the dual affinity of Ti and antibody to achieve efficient enrichment of exosomes, in addition to the mild elution property of BI-NPs due to the presence of the host-guest system. BI-NPs offer a novel approach for the isolation of exosomes, in order to further promote the application of exosomes in the required fields.
外泌体的研究在疾病诊断和预后领域具有重要意义，由于其在体液中的浓度极低，快速且低损耗地分离外泌体成为关键步骤。在此，通过主客体相互作用将钛和抗体与磁性纳米颗粒结合，成功构建了双功能免疫亲和磁性纳米颗粒（BI-NPs）。细胞培养上清液中的外泌体可被BI-NPs有效捕获，然后用α-环糊精和10%氨水溶液温和洗脱。洗脱后，捕获的外泌体蛋白含量测定高达97μg/mL。通过BI-NPs分离的外泌体中共鉴定出2822个蛋白质组，其中1060个蛋白质组来源于外泌体。同时，BI-NPs获得的外泌体蛋白质量是超速离心法分离的2倍以上，上述结果表明BI-NPs具有高纯度富集性能。这归因于BI-NPs结合了钛和抗体的双重亲和力以实现外泌体的高效富集，此外由于主客体系统的存在，BI-NPs具有温和的洗脱特性。BI-NPs为外泌体的分离提供了一种新方法，以进一步促进外泌体在所需领域的应用。
