An ELISA method for the detection and quantification of human heparanase

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• 作者：Itay Shafat ; Eyal Zcharia ; Benjamin Nisman ; Yona Nadir ; Farid Nakhoul ; Israel Vlodavsky and Neta Ilan
• 关键词：Heparanase ; ELISA ; Antibody ; Urine
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2006
• 出版时间：24 March 2006
• 年：2006
• 卷：341
• 期：4
• 页码：958-963
• 全文大小：255 K

文摘

Heparanase is a mammalian endo-β-d-glucuronidase that cleaves heparan sulfate side chains at a limited number of sites. Heparanase enzymatic activity is thought to participate in degradation and remodeling of the extracellular matrix and to facilitate cell invasion associated with tumor metastasis, angiogenesis, and inflammation. Traditionally, heparanase activity was well correlated with the metastatic potential of a large number of tumor-derived cell types. More recently, heparanase upregulation was detected in an increasing number of primary human tumors, correlating, in some cases, with poor postoperative survival and increased tumor vascularity. The present study was undertaken to develop a highly sensitive ELISA suitable for the determination and quantification of human heparanase in tissue extracts and body fluids. The assay preferentially detects the 8 + 50 kDa active heparanase heterodimer vs. the latent 65 kDa proenzyme and correlates with immunoblot analysis of heparanase containing samples. It detects heparanase at concentrations as low as 200 pg/ml and is suitable for quantification of heparanase in tissue extracts and urine.