Probing of the expression of the low-density lipoprotein receptor in vivo using an anti-receptor monoclonal antibody.
MAC188 is a rat anti-[low-density lipoprotein (LDL) receptor] monoclonal antibody (McAb) which binds to the cell surface receptor with high affinity at physiological temperatures, even in the presence of high concentrations of the natural ligand, LDL. Binding of McAb MAC188 at 37 degrees C is followed by internalization and intracellular sequestering of the receptor, which results in the transient disappearance of the receptor from the cell surface. The high binding affinity and epitope specificity of McAb MAC188 suggested that this antibody could be used to quantify receptor expression in vivo. Mixtures of radiolabelled anti-receptor antibody and a control McAb (MAC221) were injected intravenously into rabbits, and the clearance from serum and uptake into tissues was determined. A fraction of the anti-receptor McAb was cleared rapidly from the circulation by a high-affinity and saturable (receptor-dependent) process. Receptor-dependent uptake of the anti-receptor McAb was measurable in liver, adrenal glands, kidneys, spleen, kidney, thoracic aorta and heart. It was highest in liver and adrenal glands and correlated well with the level of receptor protein and the rate of LDL transport in individual tissues. Anti-receptor McAbs such as MAC188, with suitable domain specificity and binding affinity at physiological temperatures, have important advantages over the natural ligand as tracers for the receptor in vivo, and may find widespread applications in studies of the receptor status (activity) in animals and man.
Gherardi
E
E
Department of Pathology, University of Cambridge, U.K.
Bowyer
D E
DE
Fitzsimmons
C
C
Le Cras
T
T
Hutchings
A
A
Butcher
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000936:Antigen-Antibody Complex / Q000032:analysis; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood / Q000378:metabolism; D008297:Male; D008657:Metabolic Clearance Rate; D009928:Organ Specificity; D011817:Rabbits; D051381:Rats; D011973:Receptors, LDL / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D013395:Sucrose / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D013395:Sucrose