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.