Modulation of nonspecific binding in ultrafiltration protein binding studies
The aim of this study was to reduce or prevent nonspecific binding (NSB) of compounds to ultrafiltration (UF) protein binding (PB) testing units. UF units (regenerated cellulose, MWCO 10K) were used for PB and NSB measurements with or without pretreatment with 5% tween 80 (TW 80) or 5% benzalkonium...
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| Veröffentlicht in: | Pharmaceutical research 2003-07, Vol.20 (7), p.1015-1021 |
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| creator | LEE, Kyoung-Jin MOWER, Rachel HOLLENBECK, Tom CASTELO, Jesus JOHNSON, Nikole GORDON, Perry SINKO, Patrick J HOLME, Kevin LEE, Yong-Hee |
| description | The aim of this study was to reduce or prevent nonspecific binding (NSB) of compounds to ultrafiltration (UF) protein binding (PB) testing units.
UF units (regenerated cellulose, MWCO 10K) were used for PB and NSB measurements with or without pretreatment with 5% tween 80 (TW 80) or 5% benzalkonium chloride (BAK) on the filter membrane. Dosing solutions (10 microM) in human serum and pH 7.4 phosphate-buffered saline were centrifuged at 3,000 g and room temperature after 1-h incubation in UF testing units. In parallel, a 96-well equilibrium dialyzer was used for PB and NSB measurements in equilibrium dialysis (ED) at 37 degrees C for 4 h. Samples of UF and ED were analyzed by LC/MS or LSC.
Severe NSB was observed for etoposide, hydrocortisone, propranolol, and vinblastine in UF. In contrast, TW 80 or BAK pre-treatment on the filter membrane decreased the NSB from 87-95% to 13-64% without causing a significant change in membrane integrity. When NSB was below 50% as a result of pretreating agents, PB data of marker compounds were comparable to those of ED.
The pretreated membrane with TW 80 or BAK showed significantly less NSB for compounds that had a tendency toward high membrane binding. A modified UF method with pretreatment improved the performance of UF and was able to produce comparable PB results to ED. |
| doi_str_mv | 10.1023/A:1024406221962 |
| format | Article |
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UF units (regenerated cellulose, MWCO 10K) were used for PB and NSB measurements with or without pretreatment with 5% tween 80 (TW 80) or 5% benzalkonium chloride (BAK) on the filter membrane. Dosing solutions (10 microM) in human serum and pH 7.4 phosphate-buffered saline were centrifuged at 3,000 g and room temperature after 1-h incubation in UF testing units. In parallel, a 96-well equilibrium dialyzer was used for PB and NSB measurements in equilibrium dialysis (ED) at 37 degrees C for 4 h. Samples of UF and ED were analyzed by LC/MS or LSC.
Severe NSB was observed for etoposide, hydrocortisone, propranolol, and vinblastine in UF. In contrast, TW 80 or BAK pre-treatment on the filter membrane decreased the NSB from 87-95% to 13-64% without causing a significant change in membrane integrity. When NSB was below 50% as a result of pretreating agents, PB data of marker compounds were comparable to those of ED.
The pretreated membrane with TW 80 or BAK showed significantly less NSB for compounds that had a tendency toward high membrane binding. A modified UF method with pretreatment improved the performance of UF and was able to produce comparable PB results to ED.</description><identifier>ISSN: 0724-8741</identifier><identifier>EISSN: 1573-904X</identifier><identifier>DOI: 10.1023/A:1024406221962</identifier><identifier>PMID: 12880287</identifier><identifier>CODEN: PHREEB</identifier><language>eng</language><publisher>New York, NY: Springer</publisher><subject>Biological and medical sciences ; Blood Proteins - metabolism ; Caffeine ; Cellulose ; Dialyzers ; Drug interactions ; Equilibrium ; Hemodialysis ; Humans ; Medical sciences ; Membranes ; Methods ; Nonsteroidal anti-inflammatory drugs ; Pharmaceutical Preparations - metabolism ; Plasma ; Protein Binding - physiology ; Proteins ; Ultrafiltration - methods</subject><ispartof>Pharmaceutical research, 2003-07, Vol.20 (7), p.1015-1021</ispartof><rights>2003 INIST-CNRS</rights><rights>Copyright Kluwer Academic Publishers Jul 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c356t-492659a68e832836a0f62f898daff3e681d13c5f04fb3b15770468016907e8173</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14992992$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12880287$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>LEE, Kyoung-Jin</creatorcontrib><creatorcontrib>MOWER, Rachel</creatorcontrib><creatorcontrib>HOLLENBECK, Tom</creatorcontrib><creatorcontrib>CASTELO, Jesus</creatorcontrib><creatorcontrib>JOHNSON, Nikole</creatorcontrib><creatorcontrib>GORDON, Perry</creatorcontrib><creatorcontrib>SINKO, Patrick J</creatorcontrib><creatorcontrib>HOLME, Kevin</creatorcontrib><creatorcontrib>LEE, Yong-Hee</creatorcontrib><title>Modulation of nonspecific binding in ultrafiltration protein binding studies</title><title>Pharmaceutical research</title><addtitle>Pharm Res</addtitle><description>The aim of this study was to reduce or prevent nonspecific binding (NSB) of compounds to ultrafiltration (UF) protein binding (PB) testing units.
UF units (regenerated cellulose, MWCO 10K) were used for PB and NSB measurements with or without pretreatment with 5% tween 80 (TW 80) or 5% benzalkonium chloride (BAK) on the filter membrane. Dosing solutions (10 microM) in human serum and pH 7.4 phosphate-buffered saline were centrifuged at 3,000 g and room temperature after 1-h incubation in UF testing units. In parallel, a 96-well equilibrium dialyzer was used for PB and NSB measurements in equilibrium dialysis (ED) at 37 degrees C for 4 h. Samples of UF and ED were analyzed by LC/MS or LSC.
Severe NSB was observed for etoposide, hydrocortisone, propranolol, and vinblastine in UF. In contrast, TW 80 or BAK pre-treatment on the filter membrane decreased the NSB from 87-95% to 13-64% without causing a significant change in membrane integrity. When NSB was below 50% as a result of pretreating agents, PB data of marker compounds were comparable to those of ED.
The pretreated membrane with TW 80 or BAK showed significantly less NSB for compounds that had a tendency toward high membrane binding. A modified UF method with pretreatment improved the performance of UF and was able to produce comparable PB results to ED.</description><subject>Biological and medical sciences</subject><subject>Blood Proteins - metabolism</subject><subject>Caffeine</subject><subject>Cellulose</subject><subject>Dialyzers</subject><subject>Drug interactions</subject><subject>Equilibrium</subject><subject>Hemodialysis</subject><subject>Humans</subject><subject>Medical sciences</subject><subject>Membranes</subject><subject>Methods</subject><subject>Nonsteroidal anti-inflammatory drugs</subject><subject>Pharmaceutical Preparations - metabolism</subject><subject>Plasma</subject><subject>Protein Binding - physiology</subject><subject>Proteins</subject><subject>Ultrafiltration - methods</subject><issn>0724-8741</issn><issn>1573-904X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNpd0EtLAzEQAOAgiq3VszdZBL2tJpNsHt5K8QUVLwreluxuIinbpG42B_-9UVsEYZiB4WOYGYROCb4iGOj1_CYXxjAHIIrDHpqSStBSYfa2j6ZYACulYGSCjmJcYYwlUewQTQhIiUGKKVo-hS71enTBF8EWPvi4Ma2zri0a5zvn3wvni9SPg7buO__IzRBGk_s7EsfUOROP0YHVfTQn2zpDr3e3L4uHcvl8_7iYL8uWVnwsmQJeKc2lkRQk5RpbDlYq2WlrqeGSdIS2lcXMNrTJBwnMuMSEKyyMJILO0OXv3LzHRzJxrNcutqbvtTchxVpQRhWjkOH5P7gKafB5txoAuBKCsIzOtig1a9PVm8Gt9fBZ756UwcUW6Njq3g7aty7-OaYU5KBfOJh0tg</recordid><startdate>20030701</startdate><enddate>20030701</enddate><creator>LEE, Kyoung-Jin</creator><creator>MOWER, Rachel</creator><creator>HOLLENBECK, Tom</creator><creator>CASTELO, Jesus</creator><creator>JOHNSON, Nikole</creator><creator>GORDON, Perry</creator><creator>SINKO, Patrick J</creator><creator>HOLME, Kevin</creator><creator>LEE, Yong-Hee</creator><general>Springer</general><general>Springer Nature B.V</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>3V.</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20030701</creationdate><title>Modulation of nonspecific binding in ultrafiltration protein binding studies</title><author>LEE, Kyoung-Jin ; MOWER, Rachel ; HOLLENBECK, Tom ; CASTELO, Jesus ; JOHNSON, Nikole ; GORDON, Perry ; SINKO, Patrick J ; HOLME, Kevin ; LEE, Yong-Hee</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-492659a68e832836a0f62f898daff3e681d13c5f04fb3b15770468016907e8173</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Biological and medical sciences</topic><topic>Blood Proteins - metabolism</topic><topic>Caffeine</topic><topic>Cellulose</topic><topic>Dialyzers</topic><topic>Drug interactions</topic><topic>Equilibrium</topic><topic>Hemodialysis</topic><topic>Humans</topic><topic>Medical sciences</topic><topic>Membranes</topic><topic>Methods</topic><topic>Nonsteroidal anti-inflammatory drugs</topic><topic>Pharmaceutical Preparations - metabolism</topic><topic>Plasma</topic><topic>Protein Binding - physiology</topic><topic>Proteins</topic><topic>Ultrafiltration - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>LEE, Kyoung-Jin</creatorcontrib><creatorcontrib>MOWER, Rachel</creatorcontrib><creatorcontrib>HOLLENBECK, Tom</creatorcontrib><creatorcontrib>CASTELO, Jesus</creatorcontrib><creatorcontrib>JOHNSON, Nikole</creatorcontrib><creatorcontrib>GORDON, Perry</creatorcontrib><creatorcontrib>SINKO, Patrick J</creatorcontrib><creatorcontrib>HOLME, Kevin</creatorcontrib><creatorcontrib>LEE, Yong-Hee</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Pharmaceutical research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>LEE, Kyoung-Jin</au><au>MOWER, Rachel</au><au>HOLLENBECK, Tom</au><au>CASTELO, Jesus</au><au>JOHNSON, Nikole</au><au>GORDON, Perry</au><au>SINKO, Patrick J</au><au>HOLME, Kevin</au><au>LEE, Yong-Hee</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Modulation of nonspecific binding in ultrafiltration protein binding studies</atitle><jtitle>Pharmaceutical research</jtitle><addtitle>Pharm Res</addtitle><date>2003-07-01</date><risdate>2003</risdate><volume>20</volume><issue>7</issue><spage>1015</spage><epage>1021</epage><pages>1015-1021</pages><issn>0724-8741</issn><eissn>1573-904X</eissn><coden>PHREEB</coden><abstract>The aim of this study was to reduce or prevent nonspecific binding (NSB) of compounds to ultrafiltration (UF) protein binding (PB) testing units.
UF units (regenerated cellulose, MWCO 10K) were used for PB and NSB measurements with or without pretreatment with 5% tween 80 (TW 80) or 5% benzalkonium chloride (BAK) on the filter membrane. Dosing solutions (10 microM) in human serum and pH 7.4 phosphate-buffered saline were centrifuged at 3,000 g and room temperature after 1-h incubation in UF testing units. In parallel, a 96-well equilibrium dialyzer was used for PB and NSB measurements in equilibrium dialysis (ED) at 37 degrees C for 4 h. Samples of UF and ED were analyzed by LC/MS or LSC.
Severe NSB was observed for etoposide, hydrocortisone, propranolol, and vinblastine in UF. In contrast, TW 80 or BAK pre-treatment on the filter membrane decreased the NSB from 87-95% to 13-64% without causing a significant change in membrane integrity. When NSB was below 50% as a result of pretreating agents, PB data of marker compounds were comparable to those of ED.
The pretreated membrane with TW 80 or BAK showed significantly less NSB for compounds that had a tendency toward high membrane binding. A modified UF method with pretreatment improved the performance of UF and was able to produce comparable PB results to ED.</abstract><cop>New York, NY</cop><pub>Springer</pub><pmid>12880287</pmid><doi>10.1023/A:1024406221962</doi><tpages>7</tpages></addata></record> |
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| ispartof | Pharmaceutical research, 2003-07, Vol.20 (7), p.1015-1021 |
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| language | eng |
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| subjects | Biological and medical sciences Blood Proteins - metabolism Caffeine Cellulose Dialyzers Drug interactions Equilibrium Hemodialysis Humans Medical sciences Membranes Methods Nonsteroidal anti-inflammatory drugs Pharmaceutical Preparations - metabolism Plasma Protein Binding - physiology Proteins Ultrafiltration - methods |
| title | Modulation of nonspecific binding in ultrafiltration protein binding studies |
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