Magnetic separation of CD14+ cells using antibody binding with protein A expressed on bacterial magnetic particles for generating dendritic cells.
Herein the potential of a highly efficient cell separation system using bacterial magnetic particles expressing protein A (protein A-BacMPs) was demonstrated. Protein A was expressed on BacMPs using the transmembrane proteins Mms13 and MagA as anchor molecules. The evaluations of the numbers of bound antibody molecules and binding capabilities of the protein A-BacMPs using Mms13 indicated that the antibodies were efficiently introduced into protein A-BacMPs using Mms13 in comparison to MagA. In addition, the recovery ratio of the target cells on the magnetic cell separation system was enhanced by using protein A-BacMPs with Mms13. Using positive selection against peripheral blood mononuclear cells, the CD14(+) cells were separated at a purity of more than 99% by protein A-BacMPs using Mms13. Furthermore, in the evaluation of the influence of protein A-BacMPs on the separated cells, the CD14(+) cells separated using protein A-BacMPs and were successfully differentiated into dendritic cells.
Matsunaga
Tadashi
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan. tmatsuna@cc.tuat.ac.jp
Takahashi
Masayuki
M
Yoshino
Tomoko
T
Kuhara
Motoki
M
Takeyama
Haruko
H
D001426:Bacterial Proteins / Q000737:chemistry*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D018950:Lipopolysaccharide Receptors / Q000276:immunology*; D008565:Membrane Proteins / Q000737:chemistry*; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000276:immunology*
D001426:Bacterial Proteins; D018950:Lipopolysaccharide Receptors; D008565:Membrane Proteins; C405425:MpsA protein, Magnetospirillum sp.; D013205:Staphylococcal Protein A