GPC/SEC与激光光散射联用技术在食物蛋白分析中的应用研究进展

肖娜; 佟平; 熊丽姬; 陈红兵; 高金燕

doi:10.13386/j.issn1002-0306.2014.04.022

GPC/SEC与激光光散射联用技术在食物蛋白分析中的应用研究进展

肖娜^1,2,
佟平¹,
熊丽姬^1,3,
陈红兵^1,3,
高金燕²

1. 南昌大学食品科学与技术国家重点实验室
2. 南昌大学生命科学与食品工程学院
3. 南昌大学中德联合研究院

基金项目:

国家高技术研究发展计划(863计划)(2013AA102205)；国家国际科技合作专项(2013DFG31380)；国家自然科学基金(21162019)；江西省科技计划项目(20122BBG70170-2)；

详细信息

作者简介:
肖娜 (1991-) , 女, 硕士研究生, 研究方向:食品生物技术。;

中图分类号: TS207.3;O657.3
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- 文章访问数: 141
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出版历程
- 收稿日期: 2013-07-07

Application of GPC/SEC coupled with laser light scattering on food proteins

State Key Laboratory of Food Science and Technology, Nanchang University
College of Life Sciences and Food Engineering, Nanchang University
Sino-German Joint Research Institute, Nanchang University

摘要

摘要: GPC/SEC与(LLS)联用技术在分离和测定大分子物质的分子量及其分布中发挥着越来越重要的作用。该方法所需样品少、快速、准确,而且可以同时测定出物质的重均分子量(Mw)、数均分子量(Mn)、Z均分子量(Mz)、Mw/Mn、第二维里系数(A2)以及均方根半径(Rg)。目前GPC/SEC与激光光散射联用技术主要用于测定蛋白质的分子量及其分布以及研究蛋白质的聚合状态,显示了其与众不同的性能,而且该技术在食物蛋白中的应用仍值得进一步研究。
- GPC/SEC /
- 激光光散射 /
- 分子量 /
- 食物蛋白
Abstract: GPC/SEC and laser light scattering (LLS) combined technology play a more and more important role in the separation determination of molecular weight and its distribution of macromolecular substances. This method requires less samples, and also was fast and accurate, moreover it could also determine average molecular weight (Mw) , the number average molecular weight (Mn) , Z-average molecular weight (Mz) , Mw /Mn, the second virial coefficient (A2) and root mean square radius (Rg) of the material. GPC/SEC and laser light scattering detection application in protein was summarized. At present, the technology mainly used for the determination of protein molecular weight and its distribution as well as the study of protein aggregation state, and showed its distinctive performance, however, its application in food protein deserved further study.
- GPC/SEC /
- laser light scattering /
- molecular weight /
- food proteins

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参考文献(34)

[1]	王克夷.蛋白质的导论[M].北京:中国科学出版社, 2007:36-42.
[2]	孔毅, 吴梧桐, 吴如金.蛋白质分子量测定方法的比较[J].分析仪器, 2003 (2) :44-46.
[3]	Wyatt P J.Light scattering in the microbial world[J].Polymer Testing, 1983, 3 (4) :267-289.
[4]	Hjertberg T, Kulin L I, S觟rvik E.Laser light scattering as GPC detector[J].Polymer Testing, 1983, 3 (4) :267-289.
[5]	Capannoli G, Carinci V, D’Ascenzi S, et al.Dimensional analysis of saccharide conjugates with GPC and SEC-MALS:USA, US 08163296[P].2012-04-24.
[6]	Rdalo J B, Medrano R, Saiz E, et al.A simple method to determine unperturbed dimensions of polymers using size exclusion chromatography and multi-angle light scattering[J].Polymer, 2000, 41:1615-1620.
[7]	Hewa O G, Frank M, Peter K, et al.Molar mass characterization of cellulose acetates over a wide range of high DS by size exclusion chromatography with multi-angle laser light scattering detection[J].Carbohydrate Polymers, 2012, 88:96-102.
[8]	Xu Mei, Li Dong Sheng, LI Bin, et al.Comparative study on molecular weight of konjac glucomannan by gel permeation chromatography-laser light scattering-refractive Index and laser light-scattering methods[J].Spectroscopy, 2013, 10:685-698.
[9]	Se, Kazunori.Absolute molecular characterization of nonlinear block copolymers using GPC equipped with a low-angle laser light-scattering detector[J].Kobunshi Ronbunshu, 2012, 69 (5) :185-197.
[10]	Kima C, Beuve J S, Guilbert S.Study of chain branching in natural rubber using size exclusion chromatography coupled with a multi-angle light scattering detecto (rSEC-MALS) [J].European Polymer, 2009, 45:2249-2259.
[11]	Ritter A, Schmid M, Affolter S.Determination of molecular weights by size exclusion chromatography (SEC) -Results of round robin tests[J].Polymer Testing, 2010, 29:945-952.
[12]	Anastasios D.Applicability of the modfied universal calibration of gel permeation chromatography on proteins[J].Journal of Chromatography A, 2006, 1127:183-186.
[13]	Zainon O, Saphwan A A, Osman H.Molecular Characterisation of sago starch using gel permeation chromatography multi-angle Laser Light Scattering[J].Sains Malaysiana, 2010, 39 (6) :969-973.
[14]	Morgan J T, Michael C S, Christian S.The influence of macromolecular architecture on the critical aggregation concentration of large amphiphilic starch derivatives[J].Food Hydrocolloids, 2013, 31 (2) :365-374.
[15]	Liu Y G, Bo S Q, Zhu Y J.Determination of molecular weight and molecular sizes of polymers by high temperature gel permeation chromatography with a static and dynamic laser light scattering detector[J].Polymer, 2003, 44:7209-7220.
[16]	Oliva A, Llabrés M, JoséB.Farina Estimation of uncertainty in size exclusion chromatography with a double detection system (light scattering and refractive index) [J].Talanta, 2009, 78:781-789.
[17]	Leopold K K, Douglas M K, Archie E H.Size-exclusion chromatography-a review of calibration methodologies[J].Journal of biochemical and biophysical methods, 2004, 58:159-186.
[18]	Chen T Q, ADILBEKOV J, WANG J, et al.Study on molecular characteristics of four components contained in Hedysari Radix polysaccharide by gel permeation chromatographymultiangle laser light scattering technology (GPC-MALLS) [J].US National Library of Medicine National Institutes of Health, 2012, 37 (12) :1798-1803.
[19]	Ewa F S, Williams K R.Determination of molecular masses of proteins in solution:Implementation of an HPLC size exclusion chromatography and laser light scattering service in a core laboratory[J].Biomolecular Techniques, 1999 (10) :51-63.
[20]	Zhang G Y, Hayashi Y, Matsumoto S, et al.Molecular species of glycinin in some soybean cultivars[J].Phytochemistry, 2002, 60 (7) :675-681.
[21]	Emmanuelle B, Lei Tao, Chen Wen Chang, et al.Synthesis of Semitelechelic Maleimide Poly (PEGA) for Protein Conjugation By RAFT Polymerization[J].Biomacromolecules, 2009, 10 (7) :1777-1781.
[22]	Chen R, Ilasi N, Sekulica S S.Absolute molecular weight determination of hypromellose acetate succinate by size exclusion chromatography:Use of a multi-angle laser light scattering detector and a mixed solvent[J].Pharmaceutical and Biomedical Analysis, 2011, 56:743-748.
[23]	Fang Y Q, Zhang B, Wei Y M, et al.Effects of specific mechanical energy on soy protein aggregation during extrusion process studied by size exclusion chromatography coupled with multi-angle laser light scattering[J].Food Engineering, 2012, 115:220-225
[24]	Li X Y, Fang Y P, Saphwan, et al.Complexation of Bovine Serum Albumin and Sugar Beet Pectin:Structural Transitions and Phase Diagram[J].Langmuir, 2012, 28 (27) :10164-10174.
[25]	Yasushi W.Characterization of the refolding and reassembly of an integral membrane protein OmpF porin by low angle laser light scattering photometry coupled with high performance gel chromatography[J].Chromatography, 2002, 961:137-146.
[26]	Matsumura Y, Lee D S, Mori T.Molecular weight distributions ofα-lactalbumin polymers formed by mammalian and microbial transglutaminases[J].Food Hydrocolloids, 2000, 14 (1) :49-59.
[27]	Hong P, Koza S, Bouvier ESP.A review size-exclusion chromatography for the analysis of protein biotherapeutics and their aggregates[J].Liquid Chromatography&Related Technologies, 2012, 35 (27) :2923-2950.
[28]	Hassan M I, Kumar V, Singh TP, et al.Purification and characterization of zinc alpha 2-glycoprotein-prolactin inducible protein complex from human seminal plasma[J].Separation Science, 2008, 31 (12) :2318-2324.
[29]	Beaulieua M, Milena C, Sylvie L, et al.The formation of heat induced protein aggregates in whey protein/pectin mixtures studied by size exclusion chromatography coupled with multiangle laser light scattering detection[J].Food Hydrocolloids, 2005, 19:803-812.
[30]	Mallika S, Zahur U H, William W Wilson.Association tendency ofβ-lactoglobulin AB purified by gel permeation chromatography as determined by dynamic light scattering under quiescent conditions[J].Food Hydrocolloids, 1996, 10 (3) :323-328.
[31]	Bateman O A, Lubsen N H, Slingsby C.Association behaviour of human beta B1-crystallin and its truncated forms[J].Experimental Eye Research, 2001, 73 (3) :321-331.
[32]	Japp V D, Jam S.Size-exclusion chromatographymultiangle laser light scattering analysis of beta-lactoglobulin and bovine serum albumin in aqueous solution with added salt[J].Chromatography A, 2000, 867 (1-2) :105-112.
[33]	Takashi M, Yasuo M, Tatsuya S.Characterization of polydihydrosilane by SEC-MALLS and viscometry[J].Polymer, 2012, 53:2973-2978.
[34]	Christensena B E, Upset A S, Beer M U, et al.Macromolecular characterisation of three barleyβ-glucan standards by size exclusion chromatography combined with light scattering and viscometry:an inter laboratory study[J].Carbohydrate Polymers, 2001, 45:11-22.