title: Enhanced cellular uptake and transport of polyclonal immunoglobulin G and fab after their cationization.
abstract: Antibodies are poorly transported across cell membranes and biological barriers in vivo. Cationization of antibody molecules by the derivatization of surface carboxyl groups generating primary amino groups could represent a strategy for intracellular antibody delivery. Before cationization of polyclonal colchicine-specific IgG and Fab, using hexamethylenediamine the isoelectric point (pl) of native IgG and Fab (nIgG and nFab) was in the range of 5.9 9.0 and 8.7-9.3, respectively. The pI of cationized IgG and Fab (cIgG and cFab) were both higher at 8.7, 10.3 and 9.5 -11, respectively. The affinity and specificity of both IgG and Fab were not modified by cationization. When HL 60 cells were incubated with the native or cationized 125I-BSA. -IgG and -Fab, the maximal cellular uptake of clgG and cFab was 3.2 and 2.4 times higher than that of nIgG and nFab at an extracellular concentration of 500 ng/ml. Results also indicated that the uptake was dose- and temperature-dependent suggesting absorptive-mediated endocytosis of cationized antibodies by HL 60 cells. Confocal microscopy analysis indicated that the cationized antibodies were present in the plasma membranes and cytoplasm of HL 60 cells. Finally, a study with bovine arterial endothelial monolayer cells showed that the transport of cIgG and cFab through the monolayer cells was 3.3- and 4.3-fold higher for 125I-cIgG and 125I-cFab than those of the corresponding native forms.
authors.affiliation: INSERM U26 and Département de Pharmacocinétique de la Faculté de Pharmacie, Paris, France.
authors.forename: G
authors.initials: G
authors.lastname: Hong
authors.forename: O
authors.initials: O
authors.lastname: Chappey
authors.forename: E
authors.initials: E
authors.lastname: Niel
authors.forename: J M
authors.initials: JM
authors.lastname: Scherrmann
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D002412:Cations; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003078:Colchicine / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D018922:HL-60 Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000378:metabolism*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism*; D007526:Isoelectric Point; D018613:Microscopy, Confocal; D010539:Permeability; D012710:Serum Albumin, Bovine / Q000378:metabolism
chemical_list: D002412:Cations; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D003078:Colchicine