COOL, SIMON MCKENZIE,NURCOMBE, VICTOR,DOMBROWSKI, CHRISTIAN
申请号：
ES09813323
公开号：
ES2537347T3
申请日:
2009.09.11
申请国别(地区):
ES
年份:
2015
代理人:
摘要：
1. An isolated or purified heparan sulfate composition in which the heparan sulfate component is at least 97% HS / BMP2 heparan sulfate which can specifically bind to SEQ ID NO: 1 or 6, or to BMP2, in which after digestion with heparin liases (heparinase, heparitinase I and II) until the end and after subjecting the resulting disaccharide fragments to strong anion exchange HPLC (SAX-HPLC), heparan sulfate HS / BMP2 has a disaccharide composition comprising: UA-GlcN 8.2% (± 3%)； UA-GlcNAc 17.5% (± 3%)； UA-GlcN, 6S 3.7% (± 3%)； UA, 2S-GlcN 4.5% (± 3%)； UA-GlcNS 16.2% (± 3%)； UA, 2S-GlcNS, 6S 0.5% (± 3%)； in which the digestion is performed by dissolving a sample of HS / BMP2 (100 μg) in 100 mM sodium acetate / 0.2 M calcium acetate (pH 7.0)； sequentially digesting the sample with heparinase (10 mU / ml) at 37 ° C for 2 hours, followed by heparitinase I (10 mU / ml) at 37 ° C for 1 hour, followed by heparitinase II (10 mU / ml) at 37 ° C for 18 hours, followed by an aliquot of each said lyase at 37 ° C for 6 hours, in which after digestion the samples are processed on a BioGel P-2 column (1 x 120 cm) equilibrated with 0.25 M NH4HCO3 , in which the peak comprising the disaccharides is lyophilized and then dissolved in the acidified water (pH 3.5 with HCl) and then passed on a ProX PA-1 column of SAX-HPLC, in which the disaccharides elute with a linear gradient of NaCl from 0 to 1.0 M (pH, 3.5) for 60 minutes at a flow rate of 1 ml / min.1. Una composición de heparán sulfato aislada o purificada en la que el componente de heparán sulfato es al menos 97 % de heparán sulfato HS/BMP2 que puede unirse específicamente a la SEC ID Nº: 1 o 6, o a la BMP2, en la que después de la digestión con heparín liasas (heparinasa, heparitinasa I y II) hasta el final y después de someter los fragmentos de disacárido resultantes a HPLC de intercambio aniónico fuerte (SAX-HPLC), el heparán sulfato HS/BMP2 tiene una composición de disacáridos que comprende: UA-GlcN 8,2 % (± 3 %)； UA-GlcNAc 17
