Poly(\+i\N\-i\-isopropylacrylamide)-\+i\graft\-i\-polypropylene membranes containing adsorbed antibody for cell separation

• Published in: Biomaterials 2005-04, Vol.26 (11), p.1287-1292.
• Main Authors: Okamura, Aiko, Itayagoshi, Midori, Hagiwara, Taeko, Yamaguchi, Manae, Kanamori, Toshiyuki, Shinbo, Toshio, Wang, Pi-Chao
• Format: Article
• Language: English
• ISSN: 0142-9612
• DOI: 10.1016/j.biomaterials.2004.04.028

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 37DGC. Poly(N-isopropylacrylamide) has a thermoresponsive phase transition: at 32DGC water-insoluble (hydrophobic) and water-soluble (hydrophilic) states interconvert. Adsorption of antibody onto PNIPAAm-g-PP membrane at 37DGC and its desorption at 4DGC 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 37DGC 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 4DGC 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.