Chemical engineering of cell penetrating antibodies.
Antibodies, being exquisitely specific tools in biology, are routinely used to detect and identify intra-cellular structures. However, current intra-cellular application of antibodies requires that the membrane be rendered leaky, resulting in the death of cells. Here, we present a novel method to allow antibodies to penetrate the cellular membrane of living cells without affecting cell viability. A peptide (MTS, membrane transport sequence) that facilitates transport across membranes has been site-specifically attached to antibodies. MTS-antibodies enter the living cells in culture and can be detected by immunofluorescence and ELISA after extraction. Cellular structures are visualized in living cells using a specific MTS-antibody. Antibodies with membrane penetrating properties can become an important tool for the study of intra-cellular processes in living cells. Furthermore, such membrane penetrating antibodies can be used to selectively stimulate or suppress functions of the cellular machinery.
Zhao
Y
Y
Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40536, USA.
Lou
D
D
Burkett
J
J
Kohler
H
H
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000912:Antibodies, Neoplasm / Q000378:metabolism*; D003432:Cross-Linking Reagents; D003599:Cytoskeleton; D051519:Fibroblast Growth Factor 4; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D005453:Fluorescence; D006801:Humans; D016393:Lymphoma, B-Cell / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D015202:Protein Engineering; D021382:Protein Sorting Signals; D021381:Protein Transport; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D003432:Cross-Linking Reagents; C496445:FGF4 protein, human; C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D021382:Protein Sorting Signals; D011518:Proto-Oncogene Proteins; D005346:Fibroblast Growth Factors