拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器的构建与性能研究
通过将两根柱端刻蚀的毛细管插入一小段聚砜中空纤维膜中,两根毛细管在膜内距离为零,构建了拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器,研究了其衍生化性能。拼接式微膜反应器中,衍生试剂通过两根拼接毛细管之间的空隙进入反应池和目标物反应。研究表明,与膜长0.3~1.0 mm非拼接式微膜反应器相比,拼接式柱后衍生微膜反应器可保证衍生化试剂和分析物充分反应,有效改善柱效和检测灵敏度。考察了分离电压（5~10 k V）及衍生试剂浓度等因素对衍生化反应的影响,并对荧光强度和蛋白质样品浓度之间的关系进行了探究。当运行电压为10 k V,衍生试剂为含有1.5 mmol/L 2,3-萘二甲醛和60.0 mmol/...
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| Published in | 分析化学 Vol. 43; no. 10; pp. 1545 - 1550 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
上海市功能性材料化学重点实验室,华东理工大学,上海200237
2015
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0253-3820 |
| DOI | 10.11895/j.issn.0253-3820.140920 |
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| Abstract | 通过将两根柱端刻蚀的毛细管插入一小段聚砜中空纤维膜中,两根毛细管在膜内距离为零,构建了拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器,研究了其衍生化性能。拼接式微膜反应器中,衍生试剂通过两根拼接毛细管之间的空隙进入反应池和目标物反应。研究表明,与膜长0.3~1.0 mm非拼接式微膜反应器相比,拼接式柱后衍生微膜反应器可保证衍生化试剂和分析物充分反应,有效改善柱效和检测灵敏度。考察了分离电压（5~10 k V）及衍生试剂浓度等因素对衍生化反应的影响,并对荧光强度和蛋白质样品浓度之间的关系进行了探究。当运行电压为10 k V,衍生试剂为含有1.5 mmol/L 2,3-萘二甲醛和60.0 mmol/Lβ-巯基乙醇的100 mmol/L硼酸盐缓冲液（p H 9.5）时,在0.007~0.04 mg/m L浓度范围内,荧光强度与蛋白质浓度之间具有良好的线性关系,对标准蛋白BSA的检出限可达5.6 nmol/L。与毛细管区带电泳/紫外检测结果对比,此反应器几乎没有增加系统的死体积,柱效也无明显降低,具有良好的稳定性和重现性。 |
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| AbstractList | 通过将两根柱端刻蚀的毛细管插入一小段聚砜中空纤维膜中,两根毛细管在膜内距离为零,构建了拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器,研究了其衍生化性能。拼接式微膜反应器中,衍生试剂通过两根拼接毛细管之间的空隙进入反应池和目标物反应。研究表明,与膜长0.3~1.0 mm非拼接式微膜反应器相比,拼接式柱后衍生微膜反应器可保证衍生化试剂和分析物充分反应,有效改善柱效和检测灵敏度。考察了分离电压（5~10 k V）及衍生试剂浓度等因素对衍生化反应的影响,并对荧光强度和蛋白质样品浓度之间的关系进行了探究。当运行电压为10 k V,衍生试剂为含有1.5 mmol/L 2,3-萘二甲醛和60.0 mmol/Lβ-巯基乙醇的100 mmol/L硼酸盐缓冲液（p H 9.5）时,在0.007~0.04 mg/m L浓度范围内,荧光强度与蛋白质浓度之间具有良好的线性关系,对标准蛋白BSA的检出限可达5.6 nmol/L。与毛细管区带电泳/紫外检测结果对比,此反应器几乎没有增加系统的死体积,柱效也无明显降低,具有良好的稳定性和重现性。 通过将两根柱端刻蚀的毛细管插入一小段聚砜中空纤维膜中,两根毛细管在膜内距离为零,构建了拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器,研究了其衍生化性能.拼接式微膜反应器中,衍生试剂通过两根拼接毛细管之间的空隙进入反应池和目标物反应.研究表明,与膜长0.3 ～1.0 mm非拼接式微膜反应器相比,拼接式柱后衍生微膜反应器可保证衍生化试剂和分析物充分反应,有效改善柱效和检测灵敏度.考察了分离电压(5～10 kV)及衍生试剂浓度等因素对衍生化反应的影响,并对荧光强度和蛋白质样品浓度之间的关系进行了探究.当运行电压为10 kV,衍生试剂为含有1.5 mmol/L 2,3-萘二甲醛和60.0 mmol/Lβ-巯基乙醇的100 mmol/L硼酸盐缓冲液(pH 9.5)时,在0.007 ～ 0.04 mg/mL浓度范围内,荧光强度与蛋白质浓度之间具有良好的线性关系,对标准蛋白BSA的检出限可达5.6 nmol/L.与毛细管区带电泳/紫外检测结果对比,此反应器几乎没有增加系统的死体积,柱效也无明显降低,具有良好的稳定性和重现性. |
| Author | 张凌怡 任俊 彭丽 刘翻 张维冰 |
| AuthorAffiliation | 上海市功能性材料化学重点实验室,华东理工大学,上海200237 |
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| Author_FL | PENG Li LIU Fan ZHANG Wei-Bing REN Jun ZHANG Ling-Yi |
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| Keywords | 拼接式微膜反应器 柱后荧光衍生 Post-column fluorescence derivatization 毛细管电泳 Capillary electrophoresis Joint micro-membrane reactor 蛋白质 Protein |
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| Notes | A joint post-column micro-membrane reactor for fluorescent analysis of protein in capillary electrophoresis was constructed and evaluated by connecting two capillaries inside a piece of hollow fiber seamlessly. In the joint post-column micro-membrane reactor,the derivative reagent migrates into reaction pool through interval between two capillaries to react with target compounds. Compared with micro-membrane reactors with membrane lengths of 0. 3- 1. 0 mm,the joint micro-membrane reactor can guarantee complete derivatization and improve column efficiency and detection sensitivity. Influences of separation voltage（ 5- 10 k V） and concentration of derivative reagent on derivation and relationship between fluorescence intensity and sample concentration were investigated. When separation voltage was 10 k V and reaction reagent was 100 mmol / L borate buffer containing 1. 5 mmol / L 2,3-naphthalenedicarboxaldehyde and 60. 0 mmol / Lβ-mercaptoethano,wide linear calibration range from 0. 007 to 0. 04 mg / m L and hi |
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| PublicationTitle | 分析化学 |
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| Publisher | 上海市功能性材料化学重点实验室,华东理工大学,上海200237 |
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| SubjectTerms | 拼接式微膜反应器 柱后荧光衍生 毛细管电泳 蛋白质 |
| Title | 拼接式毛细管电泳柱后蛋白质荧光衍生微膜反应器的构建与性能研究 |
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