High-performance capillary electrophoresis of SDS-protein complexes using UV-transparent polymer networks
This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS-protein complexes. A study reve...
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Veröffentlicht in: | Analytical chemistry (Washington) 1992-11, Vol.64 (22), p.2665-2671 |
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creator | Ganzler, Katalin Greve, K. S Cohen, A. S Karger, B. L Guttman, Andras Cooke, N. C |
description | This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS-protein complexes. A study reveals such columns to yield significantly greater lifetime than cross-linked gels because of the flexibility of the noncovalently attached polymer chains. However, column lifetime was still found to be limited (approximately 20-40 injections), and detection at 214 nm was problematical because of the absorbance of polyacrylamide. UV-transparent polymer networks of dextran and PEG were substituted for polyacrylamide with successful molecular weight sieving of SDS-protein complexes at 214 nm. Due to their low to moderate viscosities, these networks could be routinely replaced leading to the possibility of hundreds of injections with a single column. Migration time reproducibilities of 0.5% RSD or less were found with replacement of the network. Using dextran, calibration plots of peak area vs concentration of standard protein were linear over the range of 0.5 microgram/mL up to at least 0.25 mg/mL. Furthermore, plasma samples could be directly utilized because of the strong solvating power of SDS. Rapid separation of protein mixtures are demonstrated with these UV-transparent polymer networks. |
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S ; Cohen, A. S ; Karger, B. L ; Guttman, Andras ; Cooke, N. C</creator><creatorcontrib>Ganzler, Katalin ; Greve, K. S ; Cohen, A. S ; Karger, B. L ; Guttman, Andras ; Cooke, N. C</creatorcontrib><description>This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS-protein complexes. A study reveals such columns to yield significantly greater lifetime than cross-linked gels because of the flexibility of the noncovalently attached polymer chains. However, column lifetime was still found to be limited (approximately 20-40 injections), and detection at 214 nm was problematical because of the absorbance of polyacrylamide. UV-transparent polymer networks of dextran and PEG were substituted for polyacrylamide with successful molecular weight sieving of SDS-protein complexes at 214 nm. Due to their low to moderate viscosities, these networks could be routinely replaced leading to the possibility of hundreds of injections with a single column. Migration time reproducibilities of 0.5% RSD or less were found with replacement of the network. Using dextran, calibration plots of peak area vs concentration of standard protein were linear over the range of 0.5 microgram/mL up to at least 0.25 mg/mL. Furthermore, plasma samples could be directly utilized because of the strong solvating power of SDS. 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S</creatorcontrib><creatorcontrib>Cohen, A. S</creatorcontrib><creatorcontrib>Karger, B. L</creatorcontrib><creatorcontrib>Guttman, Andras</creatorcontrib><creatorcontrib>Cooke, N. C</creatorcontrib><title>High-performance capillary electrophoresis of SDS-protein complexes using UV-transparent polymer networks</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS-protein complexes. A study reveals such columns to yield significantly greater lifetime than cross-linked gels because of the flexibility of the noncovalently attached polymer chains. However, column lifetime was still found to be limited (approximately 20-40 injections), and detection at 214 nm was problematical because of the absorbance of polyacrylamide. UV-transparent polymer networks of dextran and PEG were substituted for polyacrylamide with successful molecular weight sieving of SDS-protein complexes at 214 nm. Due to their low to moderate viscosities, these networks could be routinely replaced leading to the possibility of hundreds of injections with a single column. Migration time reproducibilities of 0.5% RSD or less were found with replacement of the network. Using dextran, calibration plots of peak area vs concentration of standard protein were linear over the range of 0.5 microgram/mL up to at least 0.25 mg/mL. Furthermore, plasma samples could be directly utilized because of the strong solvating power of SDS. 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Psychology</subject><subject>Polyethylene Glycols</subject><subject>Proteins</subject><subject>Proteins - analysis</subject><subject>Scientific imaging</subject><subject>Sodium Dodecyl Sulfate - analysis</subject><subject>Ultraviolet Rays</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkE1v1DAQhi1EVbaFE2ekCKFyqAzjr8R7RAVapBVf2xZuluOdtG6TONiJaP99jbJqEeJkye8zo3ceQp4zeMOAs7fWAYAsLYB4RBZMcaCl1vwxWeR_QXkF8ITspXQFwBiwcpfsMq5lnl0Qf-IvLumAsQmxs73DwtnBt62NtwW26MYYhssQMflUhKZYv1_TIYYRfV-40A0t3mAqpuT7i-LsnI7R9mmwEfuxGEJ722Esehx_h3idnpKdxrYJn23ffXL28cPp0QldfTn-dPRuRa2sypFKq7Apl9WmbpzgNVNgmUTHudIWS20rQMVqIRqpEBRo26Db6A2vS750vESxTw7mvbnnrwnTaDqfHOaTegxTMpWQWgDoDL78B7wKU-xzN8NZpSXnTGbocIZcDClFbMwQfZftGAbmj33zl_1Mv9iunOoONw_srDvnr7a5Tc62TdblfLrHpBRMiWXG6Iz5NOLNfWzjtSkrUSlz-nVtzr_rz-rH6pv5mfnXM29dejjifwXvAGGlqRI</recordid><startdate>19921115</startdate><enddate>19921115</enddate><creator>Ganzler, Katalin</creator><creator>Greve, K. 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S</au><au>Cohen, A. S</au><au>Karger, B. L</au><au>Guttman, Andras</au><au>Cooke, N. C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-performance capillary electrophoresis of SDS-protein complexes using UV-transparent polymer networks</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>1992-11-15</date><risdate>1992</risdate><volume>64</volume><issue>22</issue><spage>2665</spage><epage>2671</epage><pages>2665-2671</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS-protein complexes. A study reveals such columns to yield significantly greater lifetime than cross-linked gels because of the flexibility of the noncovalently attached polymer chains. However, column lifetime was still found to be limited (approximately 20-40 injections), and detection at 214 nm was problematical because of the absorbance of polyacrylamide. UV-transparent polymer networks of dextran and PEG were substituted for polyacrylamide with successful molecular weight sieving of SDS-protein complexes at 214 nm. Due to their low to moderate viscosities, these networks could be routinely replaced leading to the possibility of hundreds of injections with a single column. Migration time reproducibilities of 0.5% RSD or less were found with replacement of the network. Using dextran, calibration plots of peak area vs concentration of standard protein were linear over the range of 0.5 microgram/mL up to at least 0.25 mg/mL. Furthermore, plasma samples could be directly utilized because of the strong solvating power of SDS. Rapid separation of protein mixtures are demonstrated with these UV-transparent polymer networks.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>1284102</pmid><doi>10.1021/ac00046a003</doi><tpages>7</tpages></addata></record> |
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subjects | Analytical biochemistry: general aspects, technics, instrumentation Analytical, structural and metabolic biochemistry Biochemistry Biological and medical sciences Dextrans Electrophoresis Fundamental and applied biological sciences. Psychology Polyethylene Glycols Proteins Proteins - analysis Scientific imaging Sodium Dodecyl Sulfate - analysis Ultraviolet Rays |
title | High-performance capillary electrophoresis of SDS-protein complexes using UV-transparent polymer networks |
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