Abstract: Methods and compositions for enhancing the therapeutic activity of agents at low doses as a result of improved targeted drug delivery, namely precision delivery, are described. The methods and compositions can be used to decrease the effective administered amounts of active therapeutic agents via precision delivery to targets capable of mediating active transport across the biological barrier formed by the vasculature that lines blood vessels. Antibodies (and other targeting agent species) to precisely deliver an active ingredient (i.e., a therapeutic agent) to cells expressing, for example, a target protein on their cell membranes, are also described, as are methods of treating disease.

Abstract: Methods of delivering an agent in a tissue-specific manner, particularly lung tissue, by targeting a protein expressed on the endothelial cell surface, are described. The methods can be used for detecting, imaging and/or treating pathologies, as well as for diagnostics.

Abstract: Methods of delivering an agent in a tissue-specific manner, by targeting annexin A1, a derivative of annexin A1, or a binding partner of annexin A1, are described. The methods can be used for detecting, imaging and/or treating neoplasia, angiogenesis or neovasculature, as well as for diagnostics and methods of assessing treatment efficacy. Antibodies to annexin A1 are also described, as are methods screening for agents altering annexin A1 activity.

Abstract: Improved vascularization and tumor models, comprising a test animal having a dorsal skin window chamber, and an exogenous tissue sample implanted ectopically in the skin within the window chamber, are described, as are methods of using the models.

Abstract: Methods of delivering an agent in a tissue-specific manner, by targeting annexin A1, a derivative of annexin A1, or a binding partner of annexin A1, are described. The methods can be used for detecting, imaging and/or treating neoplasia, angiogenesis or neovasculature, as well as for diagnostics and methods of assessing treatment efficacy. Antibodies to annexin A1 are also described, as are methods screening for agents altering annexin A1 activity.

Abstract: Mechanisms and routes by which molecules can be delivered into cells, particularly endothelial cells, through the action of caveolae, G domains and other plasma membrane domains and components, are described, as are delivery systems comprising antibodies to caveolar proteins or receptors and also agents conjugated to the antibody or ligand, which can be used to deliver agents to, into, and/or across the endothelium, for example, for imaging purposes.

Abstract: Method of producing purified caveolae are described, in which a sample of interest comprising plasma membranes is subjected to an immunoisolation method in which the sample of interest is incubated with an antibody that is specific for caveolin and which binds to caveolin in its native (oligomeric) state, preferably for a brief time period, and caveolae that are bound to the antibody are separated from other materials in the sample of interest. Purified caveolae produced by the methods, and uses for the purified caveolae, are also described.

Abstract: Mechanisms and routes by which molecules can be delivered into cells, particularly endothelial cells, through the action of caveolae, G domains and other plasma membrane domains and components, are described, as are delivery systems comprising antibodies to caveolar proteins or receptors and also agents conjugated to the antibody or ligand, which can be used to deliver agents to, into, and/or across the endothelium.

Abstract: Methods of isolating and purifying caveolae, microdomains of GPI-anchored proteins, and membranes consisting essentially of caveolae associated with microdomains of GPI-anchored proteins from endothelial cell membranes are disclosed. The methods comprise coating a luminal surface of an endothelial cell membrane with an adherent first ionic material by perfusion from a luminal cavity adjacent to the endothelial cell membrane, forming a pellicle by contacting the first ionic material with a second ionic material, and isolating and purifying the pellicle. The pellicle is then processed to isolate the desired cellular component.

Abstract: Methods of isolating and purifying caveolae, microdomains of GPI-anchored proteins, and membranes consisting essentially of caveolae associated with microdomains of GPI-anchored proteins from endothelial cell membranes are disclosed. The methods comprise coating a luminal surface of an endothelial cell membrane with an adherent first ionic material (e.g. (cationic) colloidal silica) by perfusion from a luminal cavity adjacent to the endothelial cell membrane, forming a pellicle by contacting the first ionic material with a second ionic material, (e.g. acrylic polymer) and isolating and purifying the pellicle. The pellicle is then processed to isolate the desired cellular component.

Abstract: Methods of isolating and purifying caveolae, microdomains of GPI-anchored proteins, and membranes consisting essentially of caveolae associated with microdomains of GPI-anchored proteins from endothelial cell membranes are disclosed. The methods comprise coating a luminal surface of an endothelial cell membrane with an adherent first ionic material by perfusion from a luminal cavity adjacent to the endothelial cell membrane, forming a pellicle by contacting the first ionic material with a second ionic material, and isolating and purifying the pellicle. The pellicle is then processed to isolate the desired cellular component.

Abstract: Methods of isolating and purifying caveolae, microdomains of GPI-anchored proteins, and membranes consisting essentially of caveolae associated with microdomains of GPI-anchored proteins from endothelial cell membranes are disclosed. The methods comprise coating a luminal surface of an endothelial cell membrane with an adherent first ionic material by perfusion from a luminal cavity adjacent to the endothelial cell membrane, forming a pellicle by contacting the first ionic material with a second ionic material, and isolating and purifying the pellicle. The pellicle is then processed to isolate the desired cellular component.

Abstract: A process is disclosed which makes possible the isolation of the luminal endothelial cell membrane from associated tissue. It is particularly applicable to vasculature, but broadly is applicable to all tissue cavities which are accessible from adjacent perfusable lumens. The method involves the identification of characteristic molecules (primarily proteins and lipids) associated with the luminal surface of the any endothelial membrane in situ by utilizing a novel membrane-isolation scheme to separate the endothelium from associated tissue. The normal form of that tissue of interest and diseased and/or dysfunctional forms (such as tumors) can then be examined comparatively in order to identify proteins highly enriched and/or unique for normal and abnormal endothelia. This in turn permits the production of antibodies to the proteins of interest in order to develop probes specific for the abnormal tissue, such as vasculature of a tumor.

Abstract: The present invention provides methods for identifying molecular components (e.g. proteins and/or lipids) that are characteristic of vascular endothelia associated with a particular disease and are not also present in (i) normal (non-disease associated) vascular endothelia or (ii) non-disease associated vascular endothelia subjected to altered conditions that accompany the disease but are not unique to the disease.

Abstract: A process is disclosed which makes possible the isolation of the luminal endothelial cell membrane from associated tissue. It is particularly applicable to vasculature, but broadly is applicable to all tissue cavities which are accessible from adjacent perfusable lumens. The method involves the identification of characteristic molecules (primarily proteins and lipids) associated with the luminal surface of the any endothelial membrane in situ by utilizing a novel membrane-isolation scheme to separate the endothelium from associated tissue. In this method, the endothelial luminal plasmalemma of a given organ is coated with colloidal silica by perfusion, a pellicle is formed, the coated area of tissue is excised and the coated plasmalemma fragments are isolated from the cognate homogenate by centrifugation. The isolated plasmalemma attached to the pellicle can then be subjected to biochemical analysis to identify and catalogue molecules characteristic of the endothelial membrane.