title: Direct magnetic separation of immune cells from whole blood using bacterial magnetic particles displaying protein G.
abstract: 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.
authors.affiliation: Dept. of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takahashi
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yoshino
authors.forename: Haruko
authors.initials: H
authors.lastname: Takeyama
authors.forename: Tadashi
authors.initials: T
authors.lastname: Matsunaga
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology; D018189:Immunomagnetic Separation / Q000379:methods*; D008280:Magnetics; D042201:Magnetospirillum / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology