Bruce S Jacobson

US Patent:

52817008, Jan 25, 1994

Filed:

Aug 11, 1992

Appl. No.:

7/928912

Inventors:

Jan E. Schnitzer - San Diego CA
Bruce S. Jacobson - Amherst MA

Assignee:

The Regents of the University of California - Oakland CA

International Classification:

C07K 312
C07K 328

US Classification:

530412

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. The method is broadly applicable to any tissue of any organ which is adjacent to a perfusable luminal cavity and which has luminal endothelial membrane exposed to such luminal cavity.

US Patent:

55872972, Dec 24, 1996

Filed:

Jan 24, 1994

Appl. No.:

8/185433

Inventors:

Bruce S. Jacobson - Amherst MA
Jan E. Schnitzer - Boston MA

Assignee:

The Regents of the University of California - Oakland CA

International Classification:

C12Q 102
G01N 3348
C07K 118
C07K 130

US Classification:

435 29

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. The methods include isolating plasma membrane fractions from disease-associated vascular endothelial cells, from normal (nondisease-associated) vascular endothelial cells and from nondisease-associated vascular endothelial cells subjected to at least one of the altered conditions; purifying and resolving molecular components from these membrane fractions; comparing the components resolved from each of the foregoing vascular endothelial cell sources and identifying components that are present in disease-associated endothelial cells and absent from the normal or otherwise conditioned endothelial cells. The resulting endothelial cell membrane components that are unique to the disease state can be used for diagnostic and/or therapeutic purposes.

US Patent:

56100086, Mar 11, 1997

Filed:

Jun 5, 1995

Appl. No.:

8/461180

Inventors:

Jan E. Schnitzer - San Diego CA
Bruce S. Jacobson - Amherst MA

Assignee:

The Regents of the University of California - Oakland CA

International Classification:

C12Q 100

US Classification:

435 4

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. 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.