Blood-brain barrier transport of cationized immunoglobulin G: enhanced delivery compared to native protein.
IgG molecules are potential neuropharmaceuticals that may be used for therapeutic or diagnostic purposes. However, IgG molecules are excluded from entering brain, owing to a lack of transport of these plasma proteins through the brain capillary wall, or blood-brain barrier (BBB). The possibility of enhanced IgG delivery through the BBB by cationization of the proteins was explored in the present studies. Native bovine IgG molecules were cationized by covalent coupling of hexamethylenediamine and the isoelectric point was raised to greater than 10.7 based on isoelectric focusing studies. Native and cationized IgG molecules were radiolabeled with 125I and chloramine T. Cationized IgG, but not native IgG, was rapidly taken up by isolated bovine brain microvessels, which were used as an in vitro model system of the BBB. Cationized IgG binding was time and temperature dependent and was saturated by increasing concentrations of unlabeled cationized IgG (dissociation constant of the high-affinity binding site, 0.90 +/- 0.37 microM; Bmax, 1.4 +/- 0.4 nmol per mg of protein). In vivo studies documented enhanced brain uptake of 125I-labeled cationized IgG relative to [3H]albumin, and complete transcytosis of the 125I-labeled cationized IgG molecule through the BBB and into brain parenchyma was demonstrated by thaw-mount autoradiography of frozen sections of rat brain obtained after carotid arterial infusions of 125I-labeled cationized IgG. These studies demonstrate that cationization of IgG molecules greatly facilitates the transport of these plasma proteins through the BBB in vivo, and this process may provide a new strategy for IgG delivery through the BBB.
Triguero
D
D
Department of Medicine, University of California, Los Angeles School of Medicine 90024.
Buciak
J B
JB
Yang
J
J
Pardridge
W M
WM
D000818:Animals; D001812:Blood-Brain Barrier; D001921:Brain / Q000276:immunology / Q000378:metabolism; D002196:Capillaries / Q000502:physiology*; D002412:Cations; D002417:Cattle; D002560:Cerebrovascular Circulation; D007074:Immunoglobulin G / Q000378:metabolism*; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007700:Kinetics; D009928:Organ Specificity; D051381:Rats; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014316:Tritium
D002412:Cations; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D014316:Tritium; D012710:Serum Albumin, Bovine