Direct magnetic separation of immune cells from whole blood using bacterial magnetic particles displaying protein G
Direct magnetic separation of immune cells from whole blood using bacterial magnetic particles displaying protein G
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DOI：
10.1002/btpr.101
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发表时间：
2009
影响因子：
2.9
通讯作者：
Masayuki Takahashi;T. Yoshino;H. Takeyama;T. Matsunaga
中科院分区：
文献类型：
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作者：
Masayuki Takahashi;T. Yoshino;H. Takeyama;T. Matsunaga
从混合人群中直接分离靶细胞，如外周血、脐带血和骨髓，是一项用于各种治疗或诊断应用的基本技术。在这项研究中，我们制造了新的微粒，并使用这种微粒直接从全血中分离免疫细胞。磁性磁螺杆菌AMB-1能合成细胞内细菌磁性颗粒(BacMPs)，并通过基因融合技术与从BacMP膜中分离的锚定蛋白在BacMP表面表达G蛋白。展示蛋白G的BacMPs(Protein G-BacMPs)具有与多种抗体类型的高结合力，构建了不同版本的蛋白G-BacMPs与不同的抗CD单抗结合。因此，利用与抗CD单抗结合的蛋白G-BacMPs直接从全血中分离免疫细胞是可行的。全血细胞中B淋巴细胞(CD19+细胞)或T淋巴细胞(CD3+细胞)占全血细胞的比例小于0.05%，纯度高于96%。这一水平优于以前使用其他磁选方法的报告。本研究的结果证明了蛋白G-BacMP的实用性，该颗粒可能成为各种治疗或诊断应用的有力工具。©2009美国化学工程师学会生物技术。计划，2009年
Direct separation of target cells from mixed population, such as peripheral blood, umbilical cord blood, and bone marrow, is an essential technique for various therapeutic or diagnosis applications. In this study, novel particles were fabricated, and direct magnetic separation of immune cells from whole blood using such particles was performed. The magnetotactic bacterium Magnetospirillum magneticum AMB‐1 synthesizes intracellular bacterial magnetic particles (BacMPs), and protein G was expressed on the surface of the BacMPs by gene fusion techniques with anchor proteins isolated from BacMP membrane. The BacMPs displaying protein G (protein G‐BacMPs) had high binding capabilities to a wide range of antibody types, and various versions of protein G‐BacMPs binding with different anti‐CD monoclonal antibodies were constructed. Consequently, direct magnetic separation of immune cells from whole blood using protein G‐BacMPs binding with anti‐CD monoclonal antibodies was demonstrated. B lymphocytes (CD19+ cells) or T lymphocytes (CD3+ cells), which represent less than 0.05% in whole blood cells, were successfully separated at a purity level of more than 96%. This level was superior to that from previous reports using other magnetic separation approaches. The results of this study demonstrate the utility of protein G‐BacMP and this particle may become a powerful tool for various therapeutic or diagnosis applications. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009