US 7,094,580 B2 · Methods for producing pure perlecan and other heparan sulfate proteoglycans

Abstract

A method of perlecan isolation (from the EHS tumor) which produces “clean” (i.e. substantially “pure”) perlecan is disclosed. Clean perlecan is thus produced in sufficient quantities for use in a number of different in vitro and in vivo assays. In addition, this isolation method exploits a newly discovered aggregating property of a ˜220 kDa heparan sulfate proteoglycan (HSPG) observed during gel filtration chromatography, which allows it to be effectively separated from non-aggregating perlecan. The method employs specific cation exchange, anion exchange, molecular sieve chromatography and immobilized GAG affinity chromatography. It is demonstrated that there are no other contaminating proteins in the perlecan and HSPG preparations, and that the perlecan core protein is intact. Improved, clean perlecan based, rodent models of fibrillar amyloid protein deposition, accumulation and/or persistence in tissues are disclosed.

Claims13

Dependent claims

A method of preparation of substantially pure perlecan from an extract source, the method comprising the following steps:

a) isolation of an extracted perlecan by a first molecular sieve column chromatography, followed by b) running anion exchange column chromatography, followed by c) running a second molecular sieve column chromatography employing a Sephacryl S-1000 column.

Dependencies are inferred from the claim text.

Facts

Application

10/323,323Filed 18 Dec 2002 · continuation

Priority

6 Mar 1997

Published

US 2003/0153734 A114 Aug 2003

Granted

22 Aug 2006

Term ends

Not estimatedThe source records don’t support an estimate.

Classification13 codes

A61K 49/0004
Preparations for testing in vivo: screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance,…
A01K 67/027
Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals: new or modified breeds of vertebrates
A61K 49/006
Preparation for luminescence or biological staining: biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered…
A61K 51/1018
Antibodies or immunoglobulins; Fragments thereof , the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised…

Family

Application60/038,613Us provisional application · filed 6 Mar 1997

Application09/036,492Parent of this continuation · filed 6 Mar 1998

Application09/698,518Parent of this continuation · filed 26 Oct 2000

US 2003/0153734 A1Pre-grant publication · published 14 Aug 2003

US 7,094,580 B2This patent

From the US filing history. Foreign family members aren’t listed.

Methods for producing pure perlecan and other heparan sulfate proteoglycans

Life of this patent

Methods for producing pure perlecan and other heparan sulfate proteoglycans

Life of this patent

Abstract

Claims13