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Carbohydrate-Based Probes for Detection of Cellular Lectins
Carbohydrate (spacered saccharide residue, Glyc) probes with various tags were synthesized as analytical tools for study of cellular lectins, i.e., Glyc–polyacrylamide–3H, Glyc–PAA–biotin, Glyc–PAA–fluorescein (flu), and Glyc–PAA–digoxigenin, where PAA is a soluble polyacrylamide carrier of ≈30 kDa....
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Published in: | Analytical biochemistry 1998-12, Vol.265 (2), p.282-289 |
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creator | Galanina, Oxana E. Tuzikov, Alexander B. Rapoport, Evgenia Le Pendu, Jacques Bovin, Nicolai V. |
description | Carbohydrate (spacered saccharide residue, Glyc) probes with various tags were synthesized as analytical tools for study of cellular lectins, i.e., Glyc–polyacrylamide–3H, Glyc–PAA–biotin, Glyc–PAA–fluorescein (flu), and Glyc–PAA–digoxigenin, where PAA is a soluble polyacrylamide carrier of ≈30 kDa. Binding of all types of probes, where Glyc is the sialyl Lewis X (SiaLeX) tetrasaccharide or a blank saccharide, was assessed using Chinese hamster ovary (CHO) cells either transfected with the E-selectin cDNA or mock-transfected. High binding of SiaLeX–PAA–3H to E-selectin-transfected cells and absence of binding to control cells (both native and permeabilized) allowed the conclusion that the polyacrylamide carrier and the spacer arm do not contribute significantly to the binding. The biotinylated probe showed a high level of nonspecific binding in cell enzyme-linked assays. A similarly built digoxigenin-labeled probe was significantly better. In flow cytometry assays, the fluorescein probe demonstrated a specific binding to E-selectin-transfected cells of a similar level to that given by an anti-E-selectin antibody. In addition, it could be inhibited by the anti-E-selectin antibody, further demonstrating specificity. Tumors were obtained from nude mice by injection of CHO E-selectin or mock-transfected cells. The fluorescent SiaLeX–PAA–flu probe could bind to tumor sections from E-selectin-positive CHO cells, but not from control CHO cells. These probes can thus be used to reveal specifically complex carbohydrate-binding sites on cells either in culture or on tissue sections. |
doi_str_mv | 10.1006/abio.1998.2859 |
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Binding of all types of probes, where Glyc is the sialyl Lewis X (SiaLeX) tetrasaccharide or a blank saccharide, was assessed using Chinese hamster ovary (CHO) cells either transfected with the E-selectin cDNA or mock-transfected. High binding of SiaLeX–PAA–3H to E-selectin-transfected cells and absence of binding to control cells (both native and permeabilized) allowed the conclusion that the polyacrylamide carrier and the spacer arm do not contribute significantly to the binding. The biotinylated probe showed a high level of nonspecific binding in cell enzyme-linked assays. A similarly built digoxigenin-labeled probe was significantly better. In flow cytometry assays, the fluorescein probe demonstrated a specific binding to E-selectin-transfected cells of a similar level to that given by an anti-E-selectin antibody. In addition, it could be inhibited by the anti-E-selectin antibody, further demonstrating specificity. Tumors were obtained from nude mice by injection of CHO E-selectin or mock-transfected cells. The fluorescent SiaLeX–PAA–flu probe could bind to tumor sections from E-selectin-positive CHO cells, but not from control CHO cells. These probes can thus be used to reveal specifically complex carbohydrate-binding sites on cells either in culture or on tissue sections.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1006/abio.1998.2859</identifier><identifier>PMID: 9882404</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Biotin - chemistry ; Carbohydrates - chemistry ; CHO Cells ; Cricetinae ; Digoxigenin - chemistry ; DNA, Complementary ; E-Selectin - analysis ; E-Selectin - genetics ; Fluorescein - chemistry ; Mice ; Molecular Probes</subject><ispartof>Analytical biochemistry, 1998-12, Vol.265 (2), p.282-289</ispartof><rights>1998 Academic Press</rights><rights>Copyright 1998 Academic Press.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-7ab49261cc8c35a1ddda0a4703584f519b9a7f052d9e6319a09f66f6ea6183663</citedby><cites>FETCH-LOGICAL-c339t-7ab49261cc8c35a1ddda0a4703584f519b9a7f052d9e6319a09f66f6ea6183663</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9882404$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Galanina, Oxana E.</creatorcontrib><creatorcontrib>Tuzikov, Alexander B.</creatorcontrib><creatorcontrib>Rapoport, Evgenia</creatorcontrib><creatorcontrib>Le Pendu, Jacques</creatorcontrib><creatorcontrib>Bovin, Nicolai V.</creatorcontrib><title>Carbohydrate-Based Probes for Detection of Cellular Lectins</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Carbohydrate (spacered saccharide residue, Glyc) probes with various tags were synthesized as analytical tools for study of cellular lectins, i.e., Glyc–polyacrylamide–3H, Glyc–PAA–biotin, Glyc–PAA–fluorescein (flu), and Glyc–PAA–digoxigenin, where PAA is a soluble polyacrylamide carrier of ≈30 kDa. Binding of all types of probes, where Glyc is the sialyl Lewis X (SiaLeX) tetrasaccharide or a blank saccharide, was assessed using Chinese hamster ovary (CHO) cells either transfected with the E-selectin cDNA or mock-transfected. High binding of SiaLeX–PAA–3H to E-selectin-transfected cells and absence of binding to control cells (both native and permeabilized) allowed the conclusion that the polyacrylamide carrier and the spacer arm do not contribute significantly to the binding. The biotinylated probe showed a high level of nonspecific binding in cell enzyme-linked assays. A similarly built digoxigenin-labeled probe was significantly better. In flow cytometry assays, the fluorescein probe demonstrated a specific binding to E-selectin-transfected cells of a similar level to that given by an anti-E-selectin antibody. In addition, it could be inhibited by the anti-E-selectin antibody, further demonstrating specificity. Tumors were obtained from nude mice by injection of CHO E-selectin or mock-transfected cells. The fluorescent SiaLeX–PAA–flu probe could bind to tumor sections from E-selectin-positive CHO cells, but not from control CHO cells. These probes can thus be used to reveal specifically complex carbohydrate-binding sites on cells either in culture or on tissue sections.</description><subject>Animals</subject><subject>Biotin - chemistry</subject><subject>Carbohydrates - chemistry</subject><subject>CHO Cells</subject><subject>Cricetinae</subject><subject>Digoxigenin - chemistry</subject><subject>DNA, Complementary</subject><subject>E-Selectin - analysis</subject><subject>E-Selectin - genetics</subject><subject>Fluorescein - chemistry</subject><subject>Mice</subject><subject>Molecular Probes</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><recordid>eNp1kM1LxDAQxYMo67p69Sb05K110rRpgyetn7CgBz2HNJlgpNto0gr739uyizdPA_PePOb9CDmnkFEAfqVa5zMqRJ3ldSkOyJKC4CkwEIdkCQAszbmojslJjJ8AlBYlX5CFqOu8gGJJrhsVWv-xNUENmN6qiCZ5Db7FmFgfkjscUA_O94m3SYNdN3YqJOt518dTcmRVF_FsP1fk_eH-rXlK1y-Pz83NOtWMiSGtVFuInFOta81KRY0xClRRASvrwpZUtEJVFsrcCOSMCgXCcm45Kk5rxjlbkctd7lfw3yPGQW5c1NMzqkc_RskFZUXJZmO2M-rgYwxo5VdwGxW2koKcacmZlpxpyZnWdHCxTx7bDZo_-x7PpNc7Had6Pw6DjNphr9G4MDGQxrv_on8BS1Z3xg</recordid><startdate>19981215</startdate><enddate>19981215</enddate><creator>Galanina, Oxana E.</creator><creator>Tuzikov, Alexander B.</creator><creator>Rapoport, Evgenia</creator><creator>Le Pendu, Jacques</creator><creator>Bovin, Nicolai V.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19981215</creationdate><title>Carbohydrate-Based Probes for Detection of Cellular Lectins</title><author>Galanina, Oxana E. ; Tuzikov, Alexander B. ; Rapoport, Evgenia ; Le Pendu, Jacques ; Bovin, Nicolai V.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-7ab49261cc8c35a1ddda0a4703584f519b9a7f052d9e6319a09f66f6ea6183663</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Animals</topic><topic>Biotin - chemistry</topic><topic>Carbohydrates - chemistry</topic><topic>CHO Cells</topic><topic>Cricetinae</topic><topic>Digoxigenin - chemistry</topic><topic>DNA, Complementary</topic><topic>E-Selectin - analysis</topic><topic>E-Selectin - genetics</topic><topic>Fluorescein - chemistry</topic><topic>Mice</topic><topic>Molecular Probes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Galanina, Oxana E.</creatorcontrib><creatorcontrib>Tuzikov, Alexander B.</creatorcontrib><creatorcontrib>Rapoport, Evgenia</creatorcontrib><creatorcontrib>Le Pendu, Jacques</creatorcontrib><creatorcontrib>Bovin, Nicolai V.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Galanina, Oxana E.</au><au>Tuzikov, Alexander B.</au><au>Rapoport, Evgenia</au><au>Le Pendu, Jacques</au><au>Bovin, Nicolai V.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Carbohydrate-Based Probes for Detection of Cellular Lectins</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1998-12-15</date><risdate>1998</risdate><volume>265</volume><issue>2</issue><spage>282</spage><epage>289</epage><pages>282-289</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>Carbohydrate (spacered saccharide residue, Glyc) probes with various tags were synthesized as analytical tools for study of cellular lectins, i.e., Glyc–polyacrylamide–3H, Glyc–PAA–biotin, Glyc–PAA–fluorescein (flu), and Glyc–PAA–digoxigenin, where PAA is a soluble polyacrylamide carrier of ≈30 kDa. Binding of all types of probes, where Glyc is the sialyl Lewis X (SiaLeX) tetrasaccharide or a blank saccharide, was assessed using Chinese hamster ovary (CHO) cells either transfected with the E-selectin cDNA or mock-transfected. High binding of SiaLeX–PAA–3H to E-selectin-transfected cells and absence of binding to control cells (both native and permeabilized) allowed the conclusion that the polyacrylamide carrier and the spacer arm do not contribute significantly to the binding. The biotinylated probe showed a high level of nonspecific binding in cell enzyme-linked assays. A similarly built digoxigenin-labeled probe was significantly better. In flow cytometry assays, the fluorescein probe demonstrated a specific binding to E-selectin-transfected cells of a similar level to that given by an anti-E-selectin antibody. In addition, it could be inhibited by the anti-E-selectin antibody, further demonstrating specificity. Tumors were obtained from nude mice by injection of CHO E-selectin or mock-transfected cells. The fluorescent SiaLeX–PAA–flu probe could bind to tumor sections from E-selectin-positive CHO cells, but not from control CHO cells. These probes can thus be used to reveal specifically complex carbohydrate-binding sites on cells either in culture or on tissue sections.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>9882404</pmid><doi>10.1006/abio.1998.2859</doi><tpages>8</tpages></addata></record> |
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subjects | Animals Biotin - chemistry Carbohydrates - chemistry CHO Cells Cricetinae Digoxigenin - chemistry DNA, Complementary E-Selectin - analysis E-Selectin - genetics Fluorescein - chemistry Mice Molecular Probes |
title | Carbohydrate-Based Probes for Detection of Cellular Lectins |
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