Poly(N-isopropylacrylamide)-graft-polypropylene membranes containing adsorbed antibody for cell separation.
We developed a novel selective cell-separation method based on using a poly(N-isopropylacrylamide)-graft-polypropylene (PNIPAAm-g-PP) membrane containing adsorbed monoclonal antibody specific to the target cell. This membrane was prepared by plasma-induced polymerization and soaking in an antibody solution at 37 degrees C. Poly(N-isopropylacrylamide) has a thermoresponsive phase transition: at 32 degrees C water-insoluble (hydrophobic) and water-soluble (hydrophilic) states interconvert. Adsorption of antibody onto PNIPAAm-g-PP membrane at 37 degrees C and its desorption at 4 degrees C was verified by fluorescence-microscopy of the PNIPAAm-g-PP membrane after soaking it in fluorescein-conjugated goat anti-mouse IgG in phosphate-buffered saline. PNIPAAm-g-PP membranes containing adsorbed anti-mouse CD80 monoclonal antibody preferentially captured mouse-CD80 transfected cells at 37 degrees C compared with membranes lacking antibody or containing anti-mouse CD86 monoclonal antibody. Detachment of captured cells from PNIPAAm-g-PP membranes was facilitated by washing at 4 degrees C because of the thermoresponsive phase transition of PNIPAAm. With this method, mouse CD80- or mouse CD86-transfected cells were enriched from a 1:1 cell suspension to 72% or 66%, simply and with high yield.
Okamura
Aiko
A
Research Center of Advanced Bionics, National Institute of Advanced Industrial Science and Technology, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Itayagoshi
Midori
M
Hagiwara
Taeko
T
Yamaguchi
Manae
M
Kanamori
Toshiyuki
T
Shinbo
Toshio
T
Wang
Pi-Chao
PC
D000180:Acrylic Resins / Q000737:chemistry*; D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D018122:B7-1 Antigen / Q000276:immunology; D002448:Cell Adhesion / Q000502:physiology*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D008264:Macrophages / Q000166:cytology* / Q000502:physiology*; D008422:Materials Testing; D008567:Membranes, Artificial; D051379:Mice; D011126:Polypropylenes / Q000737:chemistry*; D011485:Protein Binding
D000180:Acrylic Resins; D000906:Antibodies; D018122:B7-1 Antigen; D020099:Coated Materials, Biocompatible; D008567:Membranes, Artificial; D011126:Polypropylenes; C052970:poly-N-isopropylacrylamide