人血清转铁蛋白标准物质纯化

doi:10.12034/j.issn.1009-606X.217300

过程工程学报 ›› 2018, Vol. 18 ›› Issue (2): 387-392.DOI: 10.12034/j.issn.1009-606X.217300

人血清转铁蛋白标准物质纯化

孔金隆^1,2，全灿^2*，王启迪³，罗坚⁴，周俊波³，孙永跃¹，李增兰⁴

1. 天津理工大学化学化工学院，天津 300384；2. 中国计量科学研究院化学计量与分析科学研究所，北京 100029； 3. 北京化工大学机电工程学院，北京 100029；4. 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

收稿日期:2017-08-15 修回日期:2017-09-22 出版日期:2018-04-22 发布日期:2018-04-10
通讯作者: 孔金隆 pharmacy_kong@foxmail.com
基金资助:
国家重点研发计划”国家质量基础的共性技术研究与应用”课题;中国计量科学研究院基本科研业务费

Purification of Human Serum Transferrin Reference Material

Jinlong KONG^1,2, Can QUAN^2*, Qidi WANG³, Jian LUO⁴, Junbo ZHOU³, Yongyue SUN¹, Zenglan LI⁴

1. School of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin 300384, China; 2. Division of Metrology in Chemistry, National Institute of Metrology, Beijing 100029, China; 3. College of Mechanical and Electrical Engineering, Beijing University of Chemical Technology, Beijing 100029, China; 4. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, CAS, Beijing 100190, China

Received:2017-08-15 Revised:2017-09-22 Online:2018-04-22 Published:2018-04-10
Contact: Jin LongKONG pharmacy_kong@foxmail.com

摘要/Abstract

摘要： 建立了一种从Cohn血浆分离副产物组分IV中纯化高纯度转铁蛋白(Tf)标准物质的技术，对Cohn组分IV进行复溶、离心和过滤，采用Capto DEAE阴离子交换色谱进行初分离，再通过穿透式Octyl-Sepharose 4 Fast Flow疏水色谱进一步精制；对纯化产品进行了结构、纯度、活性等表征，采用十二烷基磺酸钠?聚丙烯酰胺凝胶电泳、基质辅助激光解吸电离飞行时间质谱、高效液相色谱及酶联免疫吸附法对Tf标准物质进行了分析. 结果表明，Tf产品纯度大于99%，收率为79%；圆二色光谱分析显示Tf的二级结构未发生改变，铁离子结合能力实验表明Tf可结合2个铁离子，保持活性.

关键词: 人血清转铁蛋白, 标准物质, Cohn组分IV, 疏水色谱, 离子交换色谱

Abstract: A two-step chromatographic method of purifying transferrin (Tf) from Cohn fraction IV, a byproduct of the Cohn plasma fractionation process, was established. After diluting, centrifugation and filtration, the Cohn fraction IV was firstly purified by Capto DEAE ion-exchange chromatography, then further purified by hydrophobic interaction chromatography. The characterization of purified Tf product was analyzed by SDS?PAGE, HPLC and ELISA. The results indicated that the purity of Tf was greater than 99% and the yield was 79%. The circular dichroism spectrum proved that the secondary structure of the purified Tf product have not changed, and iron binding ability experiment proved that the purified Tf product can bind two iron ions.

Key words: human serum transferrin, reference material, cohn fraction IV, hydrophobic interaction chromatography, ion exchange chromatography

孔金隆全灿王启迪罗坚周俊波孙永跃李增兰. 人血清转铁蛋白标准物质纯化[J]. 过程工程学报, 2018, 18(2): 387-392.

Jinlong KONG Can QUAN Qidi WANG Jian LUO Junbo ZHOU Yongyue SUN Zenglan LI. Purification of Human Serum Transferrin Reference Material[J]. Chin. J. Process Eng., 2018, 18(2): 387-392.

参考文献

[1]Crichton, R R, Charloteauxwauters M.Iron Transport and Storage[J].Eur. J. Biochem., 1987, 164(3):485-506
[2]Ciuraszkiewicz J, Olczak M, Watorek W.Isolation and Characterisation of Crocodile and Python Ovotransferrins[J].Acta. Biochim. Pol., 2007, 54(1):175-182
[3]Dietrich M A, ?mijewski D, Karol H, et al.isolation and characterization of transferrin from common carp (cyprinus carpio l) seminal plasma[J].Fish Shellfish Immun., 2010, 29(1):66-74
[4]Mudumana S P, Wan H, Singh M, et al.Expression Analyses of Zebrafish Transferrin,Ifabp,and Elastaseb Mrnas as Differentiation Markers for the Three Major Endodermal Organs: Liver,Intestine,and Exocrine Pancreas[J].Dev. Dynam., 2004, 230(1):165-173
[5]Dowling P, Palmerini V, Henry M, et al.Transferrin-bound Proteins as Potential Biomarkers for Advanced Breast Cancer Patients [J].Bba Clinical, 2014, 2(C):24-30
[6]王卓，赵雄，吕茂民，等.血液制品的现状与展望[J].生物工程学报, 2011, 27(5):730-746
[7]Li, Hongyan, Qian, Zhong Ming.Transferrintransferrin receptor-mediated drug delivery[J].Medicinal Research Reviews, 2010, 22(3):225-250
[8]Qian Z M, Li H, Sun H, et al.Targeted drug delivery via the transferrin receptor-mediated endocytosis pathway[J].Pharmacological Reviews, 2002, 54(4):561-587
[9]Daniels T R, Delgado T, Helguera G, et al.The transferrin receptor part II: targeted delivery of therapeutic agents into cancer cells[J].Clinical Immunology, 2006, 121(2):159-176
[10]Ascione E, Muscariello L, Maiello V, et al.A Simple Method for Large-scale Purification of Plasma-derived Apo-transferrin[J].Biotechnol. Appl. Bioc., 2010, 57(3):87-95
[11]Wu L, Wu J, Zhang J, et al.A Simple Method for Obtaining Transferrins from Human Plasma and Porcine Serum: Preparations and Properties[J].J. Chromatogr. B., 2008, 867(1):62-68
[12]McCann K B, Hughes B, Wu J, et al.Purification of Transferrin from Cohn Supernatant I Using Ion-exchange Chromatography[J].Biotechnol. Appl. Bioc., 2005, 42(3):211-217
[13]白高英.Cohn组分IV沉淀中血浆蛋白的分离工艺及过程自动化研究 [D]. 兰州: 兰州大学, 2010, 43-44.
[14]Sa?l?gil D, ?enel S, Yavuz H, et al.Purification of Transferrin by Magnetic Immunoaffinity Beads[J].J. Sep. Sci., 2015, 38(15):2729-2736
[15]Van Scherpenzeel M, Steenbergen G, Morava E, et al.High-resolution Mass Spectrometry Glycoprofiling of Intact Transferrin for Diagnosis and Subtype Identification in the Congenital Disorders of Glycosylation[J].Transl. Res., 2015, 166(6):639-649
[16]Ordonez Y N, Anton R F, Davis W C.Quantification of Total Serum Transferrin and Transferrin Sialoforms in Human Serum； An Alternative Method for the Determination of Carbohydrate-deficient Transferrin in Clinical Samples[J].Anal. Methods-Uk, 2014, 6(12):3967-3974
[17]Nowak P, ?piewak K, Nowak J, et al.Selective Separation of Ferric and Non-Ferric Forms of Human Transferrin by Capillary Micellar Electrokinetic Chromatography [J].J. Chromatogr. A, 2014, 1341(5):73-78
[18]Kogan A, Filatov V, Gusev N, et al.Immunological Study of Complex Formation Between Soluble Transferrin Receptor and Transferrin[J].Am. J. Hematol., 2005, 79(4):281-287
[19]Nowak P, ?piewak K, Brindell M, et al.Separation of Iron-Free and Iron-Saturated Forms of Transferrin and Lactoferrin Via Capillary Electrophoresis Performed in Fused-Silica and Neutral Capillaries[J].J. Chromatogr. A, 2013, 1321(4):127-132
[20]Parkkinen J, von Bonsdorff L, Ebeling F, et al.Function and Therapeutic Development of Apotransferrin[J].Vox. Sang., 2002, 83(s1):321-326
[21]Drug E, Fadeev L, Gozin M.The Impact of Highly Hydrophobic Material on the Structure of Transferrin and Its Ability to Bind Iron[J].Toxicol. Lett., 2011, 203(1):33-39

人血清转铁蛋白标准物质纯化

Purification of Human Serum Transferrin Reference Material

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 3

编辑推荐

Metrics

[1]	尤星力杨延丽苏志国张媛张松平. 鸡白痢沙门氏菌兔抗血清多抗IgG的纯化与稳定[J]. 过程工程学报, 2019, 19(6): 1204-1211.
[2]	罗天乐;丁虹;李京京;苏志国. 反相色谱法研究人粒细胞集落刺激因子的离子交换色谱复性和稀释复性[J]. , 2005, 5(2): 213-216.
[3]	王艳;周春山;王歆燕. 阴离子交换色谱法分离含钴溶液中锌、镉、铜等杂质[J]. , 2001, 1(4): 0-0.