title: Visualization of the membrane engineering concept: evidence for the specific orientation of electroinserted antibodies and selective binding of target analytes.
abstract: Membrane engineering is a generic methodology for increasing the selectivity of a cell biosensor against a target molecule, by electroinserting target-specific receptor-like molecules on the cell surface. Previous studies have elucidated the biochemical aspects of the interaction between various analytes (including viruses) and their homologous membrane-engineered cells. In the present study, purified anti-biotin antibodies from a rabbit antiserum along with in-house prepared biotinylated bovine serum albumin (BSA) were used as a model antibody-antigen pair of molecules for facilitating membrane engineering experiments. It was proven, with the aid of fluorescence microscopy, that (i) membrane-engineered cells incorporated the specific antibodies in the correct orientation and that (ii) the inserted antibodies are selectively interacting with the homologous target molecules. This is the first time the actual working concept of membrane engineering has been visualized, thus providing a final proof of the concept behind this innovative process. In addition, the fluorescence microscopy measurements were highly correlated with bioelectric measurements done with the aid of a bioelectric recognition assay.
authors.affiliation: Department of Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece.
authors.forename: Anna
authors.initials: A
authors.lastname: Kokla
authors.forename: Petros
authors.initials: P
authors.lastname: Blouchos
authors.forename: Evangelia
authors.initials: E
authors.lastname: Livaniou
authors.forename: Christos
authors.initials: C
authors.lastname: Zikos
authors.forename: Sotiris E
authors.initials: SE
authors.lastname: Kakabakos
authors.forename: Panagiota S
authors.initials: PS
authors.lastname: Petrou
authors.forename: Spyridon
authors.initials: S
authors.lastname: Kintzios
mesh_terms: D000818:Animals; D000906:Antibodies; D001698:Biomedical Engineering / Q000379:methods*; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism*; D002522:Chlorocebus aethiops; D008564:Membrane Potentials; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D014709:Vero Cells
chemical_list: D000906:Antibodies; D012710:Serum Albumin, Bovine
keywords: bioelectric recognition assay; biotinylated BSA; cell biosensors; cell membrane engineering; electroinsertion; membrane potential