Quantitative electrochemical detection of cathepsin B activity in breast cancer cell lysates using carbon nanofiber nanoelectrode arrays toward identification of cancer formation.
Quantitative electrochemical detection of cathepsin B activity in breast cancer cell lysates using carbon nanofiber nanoelectrode arrays toward identification of cancer formation.
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DOI：
10.1016/j.nano.2015.04.014
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发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Swisher LZ;Prior AM;Gunaratna MJ;Shishido S;Madiyar F;Nguyen TA;Hua DH;Li J
使用固定在由垂直排列的碳纳米纤维 (VACNF) 制成的纳米电极阵列 (NEA) 上的二茂铁 (Fc) 标记的四肽，通过交流伏安法 (ACV) 测量复杂乳腺细胞裂解物中组织蛋白酶 B 的蛋白水解活性。已经测试了四种类型的乳腺细胞，包括正常乳腺细胞（HMEC）、转化乳腺细胞（MCF-10A）、乳腺癌细胞（T47D）和转移性乳腺癌细胞（MDA-MB-231）。发现癌细胞中检测到的蛋白酶活性增加，其中 MDA-MB-231 转移癌细胞裂解物显示出最高的组织蛋白酶 B 活性。通过将重组组织蛋白酶 B 添加到免疫沉淀的 MDA-MB-231 裂解液和 HMEC 全细胞裂解液中，定量测定 MDA-MB-231 癌细胞裂解液中的等效组织蛋白酶 B 浓度。结果说明了该技术作为便携式多重电子设备的潜力，通过快速分析特定癌症相关蛋白酶的活性来进行癌症诊断和治疗监测。使用固定在由垂直排列的碳纳米纤维制成的纳米电极阵列上的二茂铁标记的四肽，通过交流伏安法测量了乳腺细胞裂解物中组织蛋白酶 B 的蛋白水解活性。已经测试了四种类型的乳腺细胞，包括正常乳腺细胞（HMEC）、转化乳腺细胞（MCF-10A）、乳腺癌细胞（T47D）和转移性乳腺癌细胞（MDA-MB-231）。发现癌细胞中检测到的蛋白酶活性增加，其中 MDA-MB-231 细胞裂解液中的值最高。该技术有望成为一种便携式多重电子设备，通过快速分析特定癌症相关蛋白酶的活性来进行癌症诊断和治疗监测。
The proteolytic activity of cathepsin B in complex breast cell lysates have been measured with alternating current voltammetry (ACV) using ferrocene (Fc)-labeled-tetrapeptides immobilized on nanoelectrode arrays (NEAs) fabricated with vertically aligned carbon nanofibers (VACNFs). Four types of breast cells have been tested, including normal breast cells (HMEC), transformed breast cells (MCF-10A), breast cancer cells (T47D), and metastatic breast cancer cells (MDA-MB-231). The detected protease activity was found increased in cancer cells, with the MDA-MB-231 metastatic cancer cell lysate showing the highest cathepsin B activity. The equivalent cathepsin B concentration in MDA-MB-231 cancer cell lysate was quantitatively determined by spiking recombinant cathepsin B into the immunoprecipitated MDA-MB-231 lysate and the HMEC whole cell lysate. The results illustrated the potential of this technique as a portable multiplex electronic device for cancer diagnosis and treatment monitoring through rapid profiling the activity of specific cancer-relevant proteases. The proteolytic activity of cathepsin B in breast cell lysates have been measured with alternating current voltammetry using ferrocene-labeled-tetrapeptides immobilized on nanoelectrode arrays fabricated with vertically aligned carbon nanofibers. Four types of breast cells have been tested, including normal breast cells (HMEC), transformed breast cells (MCF-10A), breast cancer cells (T47D), and metastatic breast cancer cells (MDA-MB-231). The detected protease activity was found increased in cancer cells, with the highest value in MDA-MB-231 cell lysate. This technique is promising as a portable multiplex electronic device for cancer diagnosis and treatment monitoring through rapid profiling the activity of specific cancer-relevant proteases.