Cloning of apoB intrabodies: Specific knockdown of apoB in HepG2 cells
Apolipoprotein (apo) B is essential for the assembly and secretion of triglyceride-rich lipoproteins made by the liver. As the sole protein component in LDL, apoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target. Single-chain antibodies (sFvs) are t...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2008-08, Vol.373 (2), p.235-240 |
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| creator | Liao, Wei Strube, Randall W. Milne, Ross W. Chen, Si-Yi Chan, Lawrence |
| description | Apolipoprotein (apo) B is essential for the assembly and secretion of triglyceride-rich lipoproteins made by the liver. As the sole protein component in LDL, apoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target. Single-chain antibodies (sFvs) are the smallest fragment of an IgG molecule capable of maintaining the antigen binding specificity of the parental antibody. In the present study, we describe the cloning and construction of two intracellular antibodies (intrabodies) to human apoB. We targeted these intrabodies to the endoplasmic reticulum for the purpose of retaining nascent apoB within the ER, thereby preventing its secretion. Expression of the 1D1 intrabody in the apoB-secreting human hepatoma cell line HepG2 resulted in marked reduction of apoB secretion. This study demonstrates the utility of an intrabody to specifically block the secretion of a protein determinant of plasma LDL as a therapeutic strategy for the treatment of hyperlipidemia. |
| doi_str_mv | 10.1016/j.bbrc.2008.06.020 |
| format | Article |
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As the sole protein component in LDL, apoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target. Single-chain antibodies (sFvs) are the smallest fragment of an IgG molecule capable of maintaining the antigen binding specificity of the parental antibody. In the present study, we describe the cloning and construction of two intracellular antibodies (intrabodies) to human apoB. We targeted these intrabodies to the endoplasmic reticulum for the purpose of retaining nascent apoB within the ER, thereby preventing its secretion. Expression of the 1D1 intrabody in the apoB-secreting human hepatoma cell line HepG2 resulted in marked reduction of apoB secretion. This study demonstrates the utility of an intrabody to specifically block the secretion of a protein determinant of plasma LDL as a therapeutic strategy for the treatment of hyperlipidemia.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2008.06.020</identifier><identifier>PMID: 18558087</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Antibodies, Monoclonal - genetics ; Antibodies, Monoclonal - immunology ; ApoB ; Apolipoproteins B - antagonists & inhibitors ; Apolipoproteins B - immunology ; Cell Line ; Cercopithecus aethiops ; Cloning, Molecular ; COS Cells ; Endoplasmic Reticulum - immunology ; HepG2 cells ; Humans ; Hybridomas ; Hyperlipoproteinemias - therapy ; Immunoglobulin Heavy Chains - genetics ; Immunoglobulin Heavy Chains - immunology ; Immunoglobulin Light Chains - immunology ; Immunoglobulin Light Chains - therapeutic use ; Immunoglobulin Variable Region - genetics ; Immunoglobulin Variable Region - immunology ; Lipoproteins ; Mice ; Single-chain antibodies ; Transfection</subject><ispartof>Biochemical and biophysical research communications, 2008-08, Vol.373 (2), p.235-240</ispartof><rights>2008 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c484t-5b35504cabc55912f581eb3dcb0285a4c279344e9c67bca57cfefabb82a458b13</citedby><cites>FETCH-LOGICAL-c484t-5b35504cabc55912f581eb3dcb0285a4c279344e9c67bca57cfefabb82a458b13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2008.06.020$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,315,782,786,887,3552,27931,27932,46002</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18558087$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liao, Wei</creatorcontrib><creatorcontrib>Strube, Randall W.</creatorcontrib><creatorcontrib>Milne, Ross W.</creatorcontrib><creatorcontrib>Chen, Si-Yi</creatorcontrib><creatorcontrib>Chan, Lawrence</creatorcontrib><title>Cloning of apoB intrabodies: Specific knockdown of apoB in HepG2 cells</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>Apolipoprotein (apo) B is essential for the assembly and secretion of triglyceride-rich lipoproteins made by the liver. As the sole protein component in LDL, apoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target. Single-chain antibodies (sFvs) are the smallest fragment of an IgG molecule capable of maintaining the antigen binding specificity of the parental antibody. In the present study, we describe the cloning and construction of two intracellular antibodies (intrabodies) to human apoB. We targeted these intrabodies to the endoplasmic reticulum for the purpose of retaining nascent apoB within the ER, thereby preventing its secretion. Expression of the 1D1 intrabody in the apoB-secreting human hepatoma cell line HepG2 resulted in marked reduction of apoB secretion. This study demonstrates the utility of an intrabody to specifically block the secretion of a protein determinant of plasma LDL as a therapeutic strategy for the treatment of hyperlipidemia.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - genetics</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>ApoB</subject><subject>Apolipoproteins B - antagonists & inhibitors</subject><subject>Apolipoproteins B - immunology</subject><subject>Cell Line</subject><subject>Cercopithecus aethiops</subject><subject>Cloning, Molecular</subject><subject>COS Cells</subject><subject>Endoplasmic Reticulum - immunology</subject><subject>HepG2 cells</subject><subject>Humans</subject><subject>Hybridomas</subject><subject>Hyperlipoproteinemias - therapy</subject><subject>Immunoglobulin Heavy Chains - genetics</subject><subject>Immunoglobulin Heavy Chains - immunology</subject><subject>Immunoglobulin Light Chains - immunology</subject><subject>Immunoglobulin Light Chains - therapeutic use</subject><subject>Immunoglobulin Variable Region - genetics</subject><subject>Immunoglobulin Variable Region - immunology</subject><subject>Lipoproteins</subject><subject>Mice</subject><subject>Single-chain antibodies</subject><subject>Transfection</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFu1DAQhi0EotvCC3BAOXFLOnZsx0YIqaxoi1SJAyBxs-zJpHibjUO8W8Tbk2hXUC5wmsN882lmfsZecKg4cH2-qUKYsBIApgJdgYBHbMXBQik4yMdsBQC6FJZ_PWGnOW8AOJfaPmUn3ChlwDQrdrnu0xCH2yJ1hR_TuyIOu8mH1EbKr4tPI2HsIhZ3Q8K7Nv0YHnDFNY1XokDq-_yMPel8n-n5sZ6xL5fvP6-vy5uPVx_WFzclSiN3pQq1UiDRB1TKctEpwynULQYQRnmJorG1lGRRNwG9arCjzodghJfKBF6fsbcH77gPW2qRlm17N05x66efLvno_u4M8Zu7TfdOSKuthlnw6iiY0vc95Z3bxryc4AdK--y0FdYIY_8LCjA1yGYxigOIU8p5ou73NhzckpPbuCUnt-TkQLs5p3no5cM7_owcg5mBNweA5m_eR5pcxkgDUhsnwp1rU_yX_xeTGKRZ</recordid><startdate>20080822</startdate><enddate>20080822</enddate><creator>Liao, Wei</creator><creator>Strube, Randall W.</creator><creator>Milne, Ross W.</creator><creator>Chen, Si-Yi</creator><creator>Chan, Lawrence</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>7T5</scope><scope>H94</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20080822</creationdate><title>Cloning of apoB intrabodies: Specific knockdown of apoB in HepG2 cells</title><author>Liao, Wei ; Strube, Randall W. ; Milne, Ross W. ; Chen, Si-Yi ; Chan, Lawrence</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c484t-5b35504cabc55912f581eb3dcb0285a4c279344e9c67bca57cfefabb82a458b13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - genetics</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>ApoB</topic><topic>Apolipoproteins B - antagonists & inhibitors</topic><topic>Apolipoproteins B - immunology</topic><topic>Cell Line</topic><topic>Cercopithecus aethiops</topic><topic>Cloning, Molecular</topic><topic>COS Cells</topic><topic>Endoplasmic Reticulum - immunology</topic><topic>HepG2 cells</topic><topic>Humans</topic><topic>Hybridomas</topic><topic>Hyperlipoproteinemias - therapy</topic><topic>Immunoglobulin Heavy Chains - genetics</topic><topic>Immunoglobulin Heavy Chains - immunology</topic><topic>Immunoglobulin Light Chains - immunology</topic><topic>Immunoglobulin Light Chains - therapeutic use</topic><topic>Immunoglobulin Variable Region - genetics</topic><topic>Immunoglobulin Variable Region - immunology</topic><topic>Lipoproteins</topic><topic>Mice</topic><topic>Single-chain antibodies</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liao, Wei</creatorcontrib><creatorcontrib>Strube, Randall W.</creatorcontrib><creatorcontrib>Milne, Ross W.</creatorcontrib><creatorcontrib>Chen, Si-Yi</creatorcontrib><creatorcontrib>Chan, Lawrence</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liao, Wei</au><au>Strube, Randall W.</au><au>Milne, Ross W.</au><au>Chen, Si-Yi</au><au>Chan, Lawrence</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning of apoB intrabodies: Specific knockdown of apoB in HepG2 cells</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2008-08-22</date><risdate>2008</risdate><volume>373</volume><issue>2</issue><spage>235</spage><epage>240</epage><pages>235-240</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>Apolipoprotein (apo) B is essential for the assembly and secretion of triglyceride-rich lipoproteins made by the liver. As the sole protein component in LDL, apoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target. Single-chain antibodies (sFvs) are the smallest fragment of an IgG molecule capable of maintaining the antigen binding specificity of the parental antibody. In the present study, we describe the cloning and construction of two intracellular antibodies (intrabodies) to human apoB. We targeted these intrabodies to the endoplasmic reticulum for the purpose of retaining nascent apoB within the ER, thereby preventing its secretion. Expression of the 1D1 intrabody in the apoB-secreting human hepatoma cell line HepG2 resulted in marked reduction of apoB secretion. This study demonstrates the utility of an intrabody to specifically block the secretion of a protein determinant of plasma LDL as a therapeutic strategy for the treatment of hyperlipidemia.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>18558087</pmid><doi>10.1016/j.bbrc.2008.06.020</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antibodies, Monoclonal - genetics Antibodies, Monoclonal - immunology ApoB Apolipoproteins B - antagonists & inhibitors Apolipoproteins B - immunology Cell Line Cercopithecus aethiops Cloning, Molecular COS Cells Endoplasmic Reticulum - immunology HepG2 cells Humans Hybridomas Hyperlipoproteinemias - therapy Immunoglobulin Heavy Chains - genetics Immunoglobulin Heavy Chains - immunology Immunoglobulin Light Chains - immunology Immunoglobulin Light Chains - therapeutic use Immunoglobulin Variable Region - genetics Immunoglobulin Variable Region - immunology Lipoproteins Mice Single-chain antibodies Transfection |
| title | Cloning of apoB intrabodies: Specific knockdown of apoB in HepG2 cells |
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