One-step separation of CD20 + cells from whole blood using bacterial magnetic particles displaying protein G
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1094-DD07-18
One-step separation of CD20+ cells from whole blood using bacterial magnetic particles displaying protein G Masayuki Takahashi, Tomoko Yoshino, Haruko Takeyama, and Tadashi Matsunaga Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan ABSTRACT Magnetic separation of target cells from mixtures, such as peripheral blood and bone marrow, has considerable practical potential in research and medical applications. Among the current cell separation techniques, magnetic cell separation using immunomagnetic particles has been routinely applied and has proven rapidness and simplicity. Magnetospirillum magneticum AMB-1 synthesizes intracellular nano-sized bacterial magnetic particles (BacMPs) that are individually enveloped by a stable lipid bilayer membrane. BacMPs, which exhibit strong ferrimagnetism, can be collected easily with commercially available permanent magnets. In this study, a novel magnetic nanoparticle displaying protein G (protein G-BacMPs) was fabricated, and one-step cell separation for direct cell separation from whole blood was performed using the
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protein G-BacMPs. B lymphocytes (CD20+ cells), which cover less than 0.3 10-2 % of whole blood cells, were separated with 93% purity using protein G-BacMPs binding with anti-CD20 monoclonal antibodies. The results of this study demonstrate the utility of protein G-BacMPs and the magnetic cell separation approach based on protein G-BacMPs in numerous applications. INTRODUCTION Cell separation is an important technology that has considerable potential in clinical and research settings. Various types of separations has been reported including hematopoietic stem cell separation for transplantation and circulating cancer cell separation from cancer patients for diagnosis. Typically, separation of target cells with high precision from whole blood is required to enrich the leukocyte numbers by density gradient centrifugation because erythrocytes constitute the vast majority of blood cells (>1000 times as many as leukocytes). However, this separation process, which is time-consuming and labor intensive, is not well adapted to automation and integrated detection. A more direct cell separation system is necessary for the separation from hazardous (diseased) blood samples, and rapid cell detection.
Among the current cell separation techniques, magnetic cell separation using immunomagnetic nanoparticles has been routinely applied and has proven rapid and simplistic compared to fluorescence-activated cell sorting system (FACS). Nano-sized magnetic particles are well suited to cell separation because they do not present the undesired effects on cells that micro-sized particles present [1]. Furthermore, the use of nano-sized magnetic particles has advantages in assay sensitivity and precision. However, the synthesis of magnetic nanoparticles that are uniform in size and shape, and that possess good dispersion properties in aqueous solutions, is challenging. Furthermore, the handling of magne
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