Methods for producing pure perlecan and other heparan sulfate proteoglycans
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United States of America Patent
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Aug 22, 2006
Grant Date -
Aug 14, 2003
app pub date -
Dec 18, 2002
filing date -
Mar 6, 1997
priority date (Note) -
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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 .sup..about.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.
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