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

Biomaterials. 2005 Apr;26(11):1287-92. doi: 10.1016/j.biomaterials.2004.04.028.

Abstract

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.

Publication types

  • Evaluation Study

MeSH terms

  • Acrylic Resins / chemistry*
  • Adsorption
  • Animals
  • Antibodies / chemistry*
  • Antibodies / immunology
  • B7-1 Antigen / immunology
  • Cell Adhesion / physiology*
  • Cell Separation / methods*
  • Cells, Cultured
  • Coated Materials, Biocompatible / chemistry
  • Macrophages / cytology*
  • Macrophages / physiology*
  • Materials Testing
  • Membranes, Artificial*
  • Mice
  • Polypropylenes / chemistry*
  • Protein Binding

Substances

  • Acrylic Resins
  • Antibodies
  • B7-1 Antigen
  • Coated Materials, Biocompatible
  • Membranes, Artificial
  • Polypropylenes
  • poly-N-isopropylacrylamide