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[1]卢圣国,段许佳,罗芳,等.硼酸亲和整体柱的制备及其在液相色谱中的应用[J].南京工业大学学报(自然科学版),2015,37(01):27-31.[doi:10.3969/j.issn.1671-7627.2015.01.005]
 LU Shengguo,DUAN Xujia,LUO Fang,et al.Preparation of boronate affinity monolithic column and its application in liquid chromatography[J].Journal of NANJING TECH UNIVERSITY(NATURAL SCIENCE EDITION),2015,37(01):27-31.[doi:10.3969/j.issn.1671-7627.2015.01.005]
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硼酸亲和整体柱的制备及其在液相色谱中的应用()
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《南京工业大学学报(自然科学版)》[ISSN:1671-7627/CN:32-1670/N]

卷:
37
期数:
2015年01期
页码:
27-31
栏目:
出版日期:
2015-01-19

文章信息/Info

Title:
Preparation of boronate affinity monolithic column and its application in liquid chromatography
文章编号:
1671-7627(2015)01-0027-05
作者:
卢圣国1段许佳1罗芳1黄和1韩毓旺2
1.南京工业大学 生物与制药工程学院,江苏 南京 210009; 2.南京工业大学 理学院,江苏 南京 210009
Author(s):
LU Shengguo1DUAN Xujia1LUO Fang1HUANG He1HAN Yuwang2
1.College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 210009,China; 2.College of Science,Nanjing Tech University,Nanjing 210009, China
关键词:
4-乙烯基苯硼酸 乙二醇二甲基丙烯酸酯 糖化血红蛋白 原位 高效液相色谱
Keywords:
vinylbenzeneboronic acid(VPBA) ethylene dimethacrylate(EDMA) glycosylated hemoglobin in situ high performance liquid chromatography(HPLC)
分类号:
O657.7
DOI:
10.3969/j.issn.1671-7627.2015.01.005
文献标志码:
A
摘要:
选用4-乙烯基苯硼酸(VPBA)为亲和单体、乙二醇/二甘醇为致孔剂,在常规液相色谱柱内,通过乙二醇二甲基丙烯酸酯(EDMA)交联作用原位聚合,制备了一种硼酸整体柱。通过优化VPBA与EDMA的质量比优化整体柱性能,并用扫描电子显微镜(SEM)及N2吸附法(BET)测定其结构及孔径分布。结果表明:聚(VPBA-co-EDMA)整体柱对辣根过氧化物酶(糖蛋白)的动态吸附量为2.338 mg/g,大于牛血清白蛋白(非糖蛋白)的吸附量0.025 mg/g。在高效液相色谱(HPLC)分离中,该柱可快速分离含顺式羟基的化合物,进一步应用于对人全血试样的糖化血红蛋白分离,表现了优良的分离性能,且准确度与酶法测试结果相近,相对标准偏差(RSD)=2.6%。
Abstract:
Poly(vinylbenzeneboronic acid-co-ethylene dimethacrylate)monolithic column was prepared by "in-situ" process with vinylbenzeneboronic acid(VPBA)as functional monomer, diethylene glycol/ ethylene glycol as porogen and ethylene dimethacrylate(EDMA)as crosslinker. Different ratios of VPBA to EDMA were used for optimizing the properties of monolithic column. The morphology and pore size distribution of the monolith were determined by scanning electron microscope(SEM)and N2 adsorption. Results showed that the dynamic adsorption capacity of horse radish peroxidase(HRP)was 2.338 mg/g, higher than that obtained for bovine serum albumin(BSA), it was 0.025 mg/g. The boronate affinity monolithic column exhibited specific capture for cis-diol-containing compounds. In addition, it could be applied to purification of glycosylated hemoglobin in the blood in high performance liquid chromatography(HPLC), thus it had good accuracy as enzymic method and the relative standard deviation(RSD)of repeatability was 2.6%.

参考文献/References:

[1] Ireland B S,Brockmeier U,Howe C M,et al.Lectin-deficient calreticulin retains full functionality as a chaperone for class I histocompatibility molecules[J].Molecular Biology of the Cell,2008,19(6):2413-2423.
[[2] Marth J D,Grewal P K.Mammalian glycosylation in immunity[J].Nature Reviews Immunology,2008,8(11):874-887.
[[3] Krishnamoorthy L,Mahal L K.Glycomic analysis:an array of technologies[J].ACS Chemical Biology,2009,4(9):715-732.
[[4] Liu X C.Boronic acids as ligands for affinity chromatography[J].Chinese Journal of Chromatography,2006,24(1):73-80.
[[5] Soundararajan S,Badawi M,Kohlrust C M,et al.Boronic acids for affinity chromatography:spectral methods for determinations of ionization and diol-binding constants[J].Analytical Biochemistry,1989,178(1):125-134.
[[6] Koyama T,Terauchi K.Synthesis and application of boronic acid-immobilized porous polymer particles:a novel packing for high-performance liquid affinity chromatography[J].Journal of Chromatography B:Biomedical Applications,1996,679(1/2):31-40.
[[7] 杨帆,毛劼,何锡文.基于巯基-烯点击反应制备有机-无机杂化硼酸亲和整体柱用于糖蛋白的选择性富集[J].色谱,2013,31(6):531-536
[[8] Yin J F,Yang G L,Wang H L,et al.Macroporous polymer monoliths fabricated by using a metal-organic coordination gel template[J].Chemical Communications,2007(44):4614-4616.
[[9] Unger K K,Skudas R,Schulte M M.Particle packed columns and monolithic columns in high-performance liquid chromatography-comparison and critical appraisal[J].Journal of Chromatography A,2008,1184(1/2):393-415.
[[10] Liu Y C,Ren L B,Liu Z.A unique boronic acid functionalized monolithic capillary for specific capture,separation and immobilization of cis-diol biomolecules[J].Chemical Communications,2011,47(17):5067-5069.
[[11] Potter O G,Breadmore M C,Hilder E F.Boronate functionalised polymer monoliths for microscale affinity chromatography[J].Analyst,2006,131(10):1094-1096.
[[12] Lin Z,Pang J,Yang H,et al.One-pot synthesis of an organic-inorganic hybrid affinity monolithic column for specific capture of glycoproteins[J].Chemical Communications,2011,47(34):9675-9677.
[[13] Krishnamurtl U,Steffes M W.Glycohemoglobin:a primary predictor of the development or reversal of complications of diabetes mellitus[J].Clinical Chemistry,2001,47(7):1157-1165.

备注/Memo

备注/Memo:
收稿日期:2013-11-07
作者简介:卢圣国(1985—),男,浙江温州人,博士生,主要研究方向为色谱填料及无机发光材料制备; 韩毓旺(联系人),教授,E-mail:hanyw@njtech.edu.cn.
引用本文:卢圣国,段许佳,罗芳,等.硼酸亲和整体柱的制备及其在液相色谱中的应用[J].南京工业大学学报:自然科学版,2015,37(1):27-31..
更新日期/Last Update: 2015-01-20