重组蛋白A免疫吸附剂的制备及吸附性能研究

杜云英1;张旭锋2;陈校园2;何小维1

北京化工大学学报(自然科学版) ›› 2011, Vol. 38 ›› Issue (3) : 92-96.

PDF(1247 KB)
欢迎访问北京化工大学学报(自然科学版),今天是 2025年7月12日 星期六
Email Alert  RSS
PDF(1247 KB)
北京化工大学学报(自然科学版) ›› 2011, Vol. 38 ›› Issue (3) : 92-96.
生物技术与环境工程

重组蛋白A免疫吸附剂的制备及吸附性能研究

  • 杜云英;张旭锋;陈校园;何小维
作者信息 +

Preparation of a recombinat protein A-based adsorbent and its adsorption properties

  • DU YunYing1;ZHANG XuFeng2;CHEN XiaoYuan2;HE XiaoWei1
Author information +
文章历史 +

摘要

以琼脂糖凝胶为载体,重组蛋白A为配基制备免疫吸附剂。过血浆动态吸附之后洗脱得到含有免疫球蛋白的洗脱液;通过分析洗脱液中免疫球蛋白的纯度可以考察吸附剂对免疫球蛋白的吸附选择性;最后利用等温静态吸附检测了吸附剂对人免疫球蛋白(hIgG)溶液的吸附行为。结果表明,洗脱液中免疫球蛋白的纯度大于92%,重组蛋白A吸附剂对免疫球蛋白吸附选择性较好;等温静态吸附结果符合Langmuir等温吸附模型;进一步分析Langmuir吸附模型,表明该吸附剂在临床治疗中有吸附量大的明显优势,重组蛋白A免疫吸附剂具有很好的临床应用前景。

Abstract

An adsorbent has been prepared by attaching recombinant protein A as the ligand to Sepharose 6FF as the support. The adsorption selectivity of the adsorbent was assessed by analyzing the abundance of eluted human immunoglobulin after dynamic adsorption of plasma. The adsorption behavior of the adsorbent was tested by isothermal adsorption of human immunoglobulin solution. It was found that the purity of human immunoglobulin in the eluant was more than 92%, suggesting that the recombinant protein A adsorbent has a strong preference for immunoglobulin. The results of isothermal adsorption are in agreement with the Langmuir isotherm equation with a dissociation constant of 0.36mg/mL and a maximum capacity of 35.7mg/mL. Our study demonstrates that the recombinant protein Abased adsorbent has good affinity for immunoglobulin and has potential clinical applications.

引用本文

导出引用
杜云英1;张旭锋2;陈校园2;何小维1
.
重组蛋白A免疫吸附剂的制备及吸附性能研究[J]. 北京化工大学学报(自然科学版), 2011, 38(3): 92-96
DU YunYing1;ZHANG XuFeng2;CHEN XiaoYuan2;HE XiaoWei1
.
Preparation of a recombinat protein A-based adsorbent and its adsorption properties[J]. Journal of Beijing University of Chemical Technology, 2011, 38(3): 92-96

参考文献

[1]Uhlen M, Guss B, Nilsson B, et al. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications[J]. J Biol Chem. 1984, 259(3): 1695-1702.
[2]Kikuchi J, Mitsui Y, Asakura T, et al. Spectroscopic investigation of tertiary fold of staphylococcal protein A to explore its engineering application[J]. Biomaterials, 1999, 20(7): 647-654.
[3]贾凌云, 杨利, 邹汉法, 等. ProteinA亲合膜色谱柱的性能及从人血浆中纯化免疫球蛋白的研究[J]. 色谱, 1998, 16(6): 16-20.
Jia L Y, Yang L, Zou H F, et al. The property of protein A affinity membrane cartridge and purification of IgG from human plasma[J]. Chinese Journal of Chromatography, 1998, 16(6): 16-20. (in Chinese)
[4]潘再法, 莫卫民, 吴仁安, 等. 蛋白A连续床亲和毛细管色谱柱的制备及性能考察[J]. 高等学校化学学报, 2002, 23(11): 2020-2072.
Pan Z F, Mo W M, Wu R A, et al. Preparation and characterization of protein A immobilized on monolithic capillary column for affinity chromatography[J]. Chemical Journal of Chinese Universites, 2002, 23(11): 2020-2072. (in Chinese)
[5]贾凌云, 杨利, 邹汉法, 等. 新型Protein A免疫吸附膜柱在血液净化中初步应用[J]. 大连理工大学学报, 2002, 42(4): 431-436.
Jia L Y, Yang L, Zou H F, et al. Primary application of novel protein A immunoadsorption membrane cartridge to blood purification[J]. Journal of Dalian Universites of Technology, 2002, 42(4): 431-436. (in Chinese)
[6]Kasper S, Neurath M F, Huber C, et al. Protein A immunoadsorption therapy for refractory, mitomycin C-associated thrombotic microangiopathy[J]. Transfusion, 2007, 47(7): 1263-1267.
[7]Poullin P, Announ N, Mugnier B, et al. Protein Aimmunoadsorption (Prosorba column) in the treatment of rheumatoid arthritis[J]. Joint Bone Spine, 2005, 72(2): 101-103.
[8]Snyder H J, Cochran S K, Balint J J, et al. Experience with protein Aimmunoadsorption in treatment-resistant adult immune thrombocytopenic purpura.[J]. Blood, 1992, 79(9): 2237-2245.
[9]陈校园. 人工重组的六联体蛋白A及其构建方法与用途, 中国: 200810028704. 4[P]. 2009-01-07.
[10]王淑兰, 梁英教. 物理化学[M]. 北京: 冶金工业出版社, 2006: 185-186.
Wang S L, Liang Y J. Physical Chemistry[M]. Beijing: Metallurgy Industry Press, 2006: 185-186. (in Chinese)
[11]Yang H, Gurgel P V, Carbonell R G. Purification of human immunoglobulin G via Fc-specific small peptide ligand affinity chromatography[J]. Journal of Chromatography A, 2009(1216): 910-918.
[12]方灿良, 赵睿, 刘阳, 等. 从组合化学肽库中筛选亲和配基[J]. 高等学校化学学报, 2003, 24(1): 52-54.
Fang C L, Zhao R, Liu Y, et al. Screening of affinity peptide ligands from combinatorial peptide libraries[J]. Chemical Journal of Chinese Universites, 2003, 24(1): 52-54. (in Chinese)
[13]Wang L, Amphlett G, Blattler W A, et al. Structural characterization of the maytansinoidmonoclonal antibody immunoconjugate, huN901DM1, by mass spectrometry[J]. Protein Sci, 2005, 14(9): 2436-2446.
[14]Schmidt E, Klinker E, Opitz A, et al. Protein A immunoadsorption: a novel and effective adjuvant treatment of severe pemphigus[J]. British Journal of Dermatology, 2003, 148(6): 1222-1229.
PDF(1247 KB)

3551

Accesses

0

Citation

Detail

段落导航
相关文章

/