无水体系中环氧氯丙烷活化琼脂糖凝胶制备高载量固定化金属亲和层析介质
Activation of Sepharose with Epichlorohydrin in Water-free System for Preparation of High Capacity Immobilized Metal Ion Affinity Adsorbent
查看参考文献16篇
文摘
|
在以二甲基亚砜为溶剂的无水体系中,利用环氧氯丙烷对琼脂糖凝胶Sepharose6Fast Flow进行活化,并偶联亚氨基二乙酸和Cu2+制备了固定化金属亲和层析介质.结果表明,该体系中环氧氯丙烷对琼脂糖凝胶的活化效率大幅度提高,在40%(φ)环氧氯丙烷,0.02g/mLNaOH及50℃,反应时间4h的优化条件下,环氧基活化密度最高达165μmol/mL,较目前报道的最高值提高50%以上.最终所制介质的Cu2+螯合密度为128.3μmol/mL,对BSA的平衡吸附容量达2.05mmol/L.以0.5mol/L咪唑为洗脱剂,被吸附的BSA洗脱率可达90%以上 |
其他语种文摘
|
Epoxy activation of agarose gel Sepharose 6 FF by epichlorohydrin (ECH) was carried out in a water-free medium with dimethyl sulfoxide as solvent. The activated agarose gel was subsequently coupled with iminodiacetic acid and Cu2+ ions for preparation of immobilized metal ions affinity chromatographic (IMAC) adsorbent. The results indicated that the epoxy activation efficiency was significantly enhanced in the water-free system, such that a epoxy density of agarose gels as high as 165 μmol/mL was achieved at the optimized conditions of 40%(ф) ECH,0.02 g/mL NaOH,50℃ and 4 h, which is about 50% higher than the highest value reported so far. The prepared IMAC adsorbent showed a high chelating capacity for Cu2+ ion up to 128.3 mol/mL, thus it ensured a high equilibrium adsorption capacity of 2.05 mmol/L for bovine serum albumin, and 90% of the adsorbed protein could be efficiently eluted when 0.5 mol/L imidazole was used as elute |
来源
|
过程工程学报
,2010,10(5):971-975 【核心库】
|
关键词
|
固定化金属亲和层析
;
琼脂糖凝胶
;
环氧氯丙烷
;
活化
|
地址
|
中国科学院过程工程研究所, 生化工程国家重点实验室, 北京, 100190
|
语种
|
中文 |
文献类型
|
研究性论文 |
ISSN
|
1009-606X |
学科
|
化学工业 |
基金
|
国家自然科学基金资助项目
;
国家863计划
;
国家973计划
|
文献收藏号
|
CSCD:4042951
|
参考文献 共
16
共1页
|
1.
Ueda E K M. Current and Prospective Applications of Metal Ion-Protein Binding.
J. Chromatogr. A,2003,988(1):1-23
|
被引
24
次
|
|
|
|
2.
Sousal F. Affinity Chromatography Approaches to Overcome the Challenges of Purifying Plasmid DNA.
Trends Biotechnol,2008,26(9):518-525
|
被引
1
次
|
|
|
|
3.
Azarkan M. Affinity Chromatography: A Useful Tool in Proteomics Studies.
J. Chromatogr. B,2007,849(1/2):81-90
|
被引
2
次
|
|
|
|
4.
Jungbauer A. Folding and Refolding of Proteins in Chromatographic Beds.
Curr. Opin. Biotechnol,2004,15(5):487-494
|
被引
11
次
|
|
|
|
5.
General Electric Healthcare Company.
GE Healthcare Data file,2009
|
被引
1
次
|
|
|
|
6.
Matsumoto I. Activation of Sepharose with Epichlorohydrin and Subsequent Immobilization of Ligand for Affinity Adsorbent.
J. Biochem,1979,85(4):1091-1098
|
被引
11
次
|
|
|
|
7.
穆成华. 肝素-琼脂糖凝胶6FF的制备及其在抗凝血酶Ⅲ分离纯化中的应用.
过程工程学报,2005,5(5):545-549
|
被引
5
次
|
|
|
|
8.
甄宇红. 环氧氯丙烷法活化琼脂糖凝胶及其动力学分析.
大连医科大学学报,2005,27(4):268-272
|
被引
5
次
|
|
|
|
9.
史清洪. 环氧氯丙烷活化琼脂糖凝胶过程强化及性能评价.
过程工程学报,2007,7(4):743-746
|
被引
13
次
|
|
|
|
10.
Scoble J A. Assay for Determining the Number of Reactive Groups on Gels Used in Affinity Chromatography and Its Application to the Optimization of the Epichlorohydrin and Divinylsulfone Activation Reactions.
J. Chromatogr. A,1996,752(1/2):67-76
|
被引
9
次
|
|
|
|
11.
Shi Q H. Chitosan-coated Silica Beads as Immobilized Metal Affinity Support for Protein Adsorption.
Biochem. Eng. J,2003,16(3):317-322
|
被引
4
次
|
|
|
|
12.
Xi F. Macroporous Chitosan Layer Coated on Non-porous Silica Gel as a Support for Metal Chelate Affinity Chromatographic Adsorbent.
J. Chromatogr. A,2004,1057(1/2):41-47
|
被引
4
次
|
|
|
|
13.
Li F. One-pot Preparation of Silica-supported Hybrid Immobilized Metal Affinity Adsorbent with Macroporous Surface Based on Surface Imprinting Coating Technique Combined with Polysaccharide Incorporated Sol-Gel Process.
J. Chromatogr. A,2006,1129(2):223-230
|
被引
10
次
|
|
|
|
14.
Chiang C L. Purification of Recombinant Enhanced Green Fluorescent Protein Expressed in Escherichia coli with New Immobilized Metal Ion Affinity Magnetic Absorbents.
J. Chromatogr. B,2008,864(1/2):116-122
|
被引
5
次
|
|
|
|
15.
Hamdane D. Expression and Purification of Untagged and Histidine-tagged Folate-dependent Trna:m5u54 Methyltransferase from Bacillus subtilis.
Protein Expr. Purif,2010,73(1):83-89
|
被引
1
次
|
|
|
|
16.
Vancan S. IMAC of Human Igg: Studies with Ida-immobilized Copper, Nickel, Zinc, and Cobalt Ions and Different Buffer Systems.
Process Biochem,2002,37(6):573-579
|
被引
2
次
|
|
|
|
|