Isolation and Detection of Exosomes Using Fe2O3 Nanoparticles
Isolation and Detection of Exosomes Using Fe2O3 Nanoparticles
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DOI：
10.1021/acsanm.0c02807
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发表时间：
2021-01-19
影响因子：
5.9
通讯作者：
Shiddiky, Muhammad J. A.
中科院分区：
文献类型：
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作者：
Farhana, Fatema Zerin;Umer, Muhammad;Shiddiky, Muhammad J. A.
具有过氧化物酶模拟活性的磁性纳米酶已被广泛研究用于分子生物传感器的开发。在此，我们报告了一种淀粉辅助的方法合成一类新的羧基功能化的氧化铁纳米粒子（C-IONP）。扫描电子显微镜和透射电子显微镜分析表明，纳米粒子具有球形形状，平均尺寸约为250 nm。通过在H2 O2存在下催化氧化3，3 '，5，5'-四甲基联苯胺（TMB）来研究C-IONP的过氧化物模拟酶活性。结果表明，纳米颗粒遵循典型的Michaelis-Menten动力学，对TMB和H2 O2表现出优异的亲和力，TMB的K-M和V-Max值分别为0.0992 mM和0.156 x 10（-8）Ms-1，H2 O2的K-M和V-Max值分别为114 mM和0.197 x 10（-8）Ms-1。C-IONP用于开发用于直接分离和定量疾病特异性外泌体的简单方法。该方法利用两步策略，其涉及（a）使用四跨膜蛋白生物标志物（即，CD 9）-官能化的C-IONP和（B）随后使用肿瘤特异性标志物（在这种情况下，卵巢癌生物标志物CA-125）电化学定量捕获的大量外来体内的疾病特异性外来体。在第一步中，C-IONP被用作“可分散的纳米载体”以捕获外泌体的大部分群体，并且在第二步中，它们被用作纳米酶以产生指示肿瘤特异性外泌体的存在的酶催化电流。计时电流分析表明，该方法对OVCAR 3细胞来源的外泌体表现出优异的特异性（线性动态范围，6.25 x 10（5）至1.0 x 10（7）外泌体/mL;检测限，1.25 x 10（6）外泌体/mL），相对标准偏差为
Magnetic nanozymes with peroxidase-mimicking activity have been widely investigated for developing molecular biosensors. Herein, we report a starch-assisted method for the synthesis of a novel class of carboxyl group-functionalized iron oxide nanoparticles (C-IONPs). Scanning electron and transmission electron microscopy analysis revealed that the nanopartides possess a spherical shape with an average size of similar to 250 nm. Peroxidase-mimicking activity of C-IONPs was investigated through catalytic oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. The results showed that nanoparticles follow typical Michaelis-Menten kinetics and exhibit excellent affinity toward TMB and H2O2 with estimated K-M and V-Max values of 0.0992 mM and 0.156 x 10(-8) Ms-1 for TMB and 114 mM and 0.197 x 10(-8) Ms-1 for H2O2, respectively. C-IONPs were used to develop a simple method for the direct isolation and quantification of disease-specific exosomes. This method utilized a two-step strategy that involved (a) initial isolation of bulk exosomes present in the sample media using tetraspanin biomarker (i.e., CD9)-functionalized C-IONPs and (b) subsequent electrochemical quantification of disease-specific exosomes within the captured bulk exosomes using tumor-specific markers (in this case, the ovarian cancer biomarker CA-125). In the first step, C-IONPs were used as "dispersible nanocarriers" to capture the bulk population of exosomes, and in the second step, they were used as nanozymes to generate an enzyme-catalyzed current indicative of the presence of tumor-specific exosomes. Chronoamperometric analysis showed that the method exhibits an excellent specificity for OVCAR3 cell-derived exosomes (linear dynamic range, 6.25 x 10(5) to 1.0 x 10(7) exosomes/mL; detection limit, 1.25 x 10(6) exosomes/mL) with a relative standard deviation of