HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS OF SDS PROTEIN COMPLEXES USING UV-TRANSPARENT POLYMER NETWORKS
HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS OF SDS PROTEIN COMPLEXES USING UV-TRANSPARENT POLYMER NETWORKS
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DOI：
10.1021/ac00046a003
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发表时间：
1992-11-15
影响因子：
7.4
通讯作者：
COOKE, NC
中科院分区：
文献类型：
--
作者：
GANZLER, K;GREVE, KS;COOKE, NC
本文展示了使用紫外透明的可替换的聚合物网络的SDS-蛋白质复合物的分子量的基础上分离。首先，线性（即非交联）聚丙烯酰胺的使用显示出提供SDS-蛋白质复合物的分子分离。一项研究表明，由于非共价连接的聚合物链的灵活性，这种柱比交联凝胶产生更长的寿命。然而，仍然发现色谱柱寿命有限（约20-40次进样），并且由于聚丙烯酰胺的吸光度，检测st 214 nm存在问题。紫外线透明的葡聚糖和PEG的聚合物网络取代聚丙烯酰胺与成功的SDS-蛋白质复合物在214 nm的分子量筛选。由于它们的低至中等粘度，这些网络可以被常规地替换，从而导致用单个柱进行数百次注射的可能性。迁移时间的重现性为0.5%RSD或更低，发现更换网络。使用葡聚糖，峰战神与标准蛋白浓度的校准图在0.5 μ g/mL至至少0.25 mg/mL的范围内呈线性。此外，由于SDS的强溶剂化能力，血浆样品可直接利用。蛋白质混合物的快速分离证明与这些UV透明的聚合物网络。
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-crossed-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 st 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 ares vs concentration of standard protein wore linear over the range of 0.5 mug/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.