六种蛋白质与生物纳米磁珠的吸附性能研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0032

生物技术通报 ›› 2020, Vol. 36 ›› Issue (6): 208-214.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0032

六种蛋白质与生物纳米磁珠的吸附性能研究

王萌慧¹, 高瑛¹, 李家璜^1,2, 华子春^1,2

1. 南京大学生命科学学院医药生物技术国家重点实验室,南京 210023;
2. 江苏省产业技术研究院医药生物技术研究所常州南京大学高新技术研究院,常州 213164

收稿日期:2020-01-08 出版日期:2020-06-26 发布日期:2020-06-28
作者简介:王萌慧,女,硕士研究生,研究方向：蛋白质结构与功能;E-mail：mhwangnju@163.com
基金资助:
国家自然科学基金项目（81573338,81630092）,国家重点研发计划干细胞及转化研究专项（2017YFA0104301）

The Study on the Adsorption Properties of Six Proteins on Biological Magnetic Nanobeads

WANG Meng-hui¹, GAO Ying¹, LI Jia-huang^1,2, HUA Zi-chun^1,2

1. School of Life Sciences,State Key Laboratory of Pharmaceutical Biotechnology,Nanjing University,Nanjing 210023;
2. Research Institute of Pharmaceutical Biotechnology,Jiangsu Industrial Technology Research Institute and High-Tech Research Institute of Nanjing University at Changzhou,Changzhou 213164

Received:2020-01-08 Published:2020-06-26 Online:2020-06-28

摘要/Abstract

摘要： 近年来,纳米磁珠在生物技术和生物医学领域受到了广泛的关注,但蛋白质与生物纳米磁珠之间往往存在非特异性吸附,故从蛋白质角度出发,研究不同蛋白质的结构及理化性质与磁珠吸附的关系。利用两种生物纳米磁珠,对6种不同蛋白质与磁珠的吸附特异性展开研究。使用蛋白质分析软件分析各个蛋白结构及蛋白质性质与磁珠吸附特异性的关系。不同的蛋白质与纳米磁珠之间存在非特异性吸附,蛋白大小与磁珠吸附蛋白的能力没有明显关联性。随着蛋白β折叠比例的降低,蛋白质与磁珠的非特异性吸附降低。β折叠比例最大的蛋白——人肿瘤坏死因子-α（TNFα）与磁珠的非特异性吸附最多,可通过吸附获得单一蛋白条带。β折叠比例最小的人膜联蛋白A5（Annexin A5）与磁珠的非特异性吸附最少。当Annexin A5融合增强绿色荧光蛋白（EGFP）后,β折叠比例升高,与磁珠的非特异性吸附增加。不同蛋白质的大小与磁珠的非特异性吸附无明显的相关性,磁珠对蛋白的非特异性吸附能力与蛋白结构的β折叠比例呈正相关,蛋白质的二级结构是影响蛋白质与磁珠非特异性吸附的重要因素。

关键词: 纳米磁珠, 不同蛋白质, 非特异性吸附, β折叠比例

Abstract: In recent years,magnetic nanobeads have received extensive attentions in the fields of biotechnology and biomedicine;however,there is often non-specific adsorption between proteins and biological magnetic nanobeads. Therefore,the relationship between the structure and physicochemical properties of proteins and magnetic bead adsorption was studied from the perspective of proteins. Using two types of biological magnetic nanobeads,the adsorption specificities of 6 different proteins on the surfaces of magnetic beads were studied. Each protein’s structure was analyzed by protein analysis software,and the structure and physicochemical property of different proteins were obtained. The relationship between protein properties and the adsorption specificity of the magnetic bead was analyzed. There was non-specific adsorption for different proteins on the surfaces of magnetic nanobeads,and the size of the protein was not significantly related to the ability of magnetic beads to adsorb proteins. As the β-sheet ratio of protein decreased,the non-specific adsorption of protein on the surfaces of magnetic beads also decreased. Human Tumor Necrosis Factor alpha（TNFα）with the largest β-sheet ratio had the most non-specific adsorption on the magnetic beads,and a single protein band was obtained by the adsorption. Human Annexin A5 with the smallest β-sheet ratio had the least non-specific adsorption on the magnetic beads. After Annexin A5 was fused with Enhanced Green Fluorescent Protein（EGFP）,the β-sheet ratio was increased,and the non-specific adsorption by magnetic beads also increased. In conclusion,there is no obvious correlation between the protein size and the non-specific adsorption on the surfaces of magnetic beads. The non-specific adsorption capacity of magnetic beads to proteins is positively related to the β-sheet ratio of the protein structure. Protein secondary structure is an important factor for the non-specific adsorption of proteins to magnetic beads.

Key words: magnetic nanobeads, different proteins, non-specific adsorption, β-sheet ratio

王萌慧, 高瑛, 李家璜, 华子春. 六种蛋白质与生物纳米磁珠的吸附性能研究[J]. 生物技术通报, 2020, 36(6): 208-214.

WANG Meng-hui, GAO Ying, LI Jia-huang, HUA Zi-chun. The Study on the Adsorption Properties of Six Proteins on Biological Magnetic Nanobeads[J]. Biotechnology Bulletin, 2020, 36(6): 208-214.

参考文献

[1] Waseem S, Ali Z, Bibi M, et al.Magnetic nanobeads:Synthesis and application in biomedicine[J]. Nanomedicine J, 2016, 3(3):147-154.
[2] Lu AH, Salabas EL, Schüth F, et al.Magnetic nanoparticles:Synthesis, protection, functionalization, and application[J]. Angew Chem Int Ed Engl, 2007, 46(8):1222-1244.
[3] 韩涛, 肖清平, 宋仲容. 磁性SiO₂微球的制备、表面修饰及其在生物医学领域的应用[J]. 国际生物医学工程杂志, 2010, 33(2):125-128.
[4] 王建忠. GST抗体包被免疫磁珠纳米佐剂的初步研究[D]. 武汉:华中农业大学, 2011.
[5] Zhang M, Zhang XH, He XW, et al.A self-assembled polydopamine film on the surface of magnetic nanoparticles for specific capture of protein[J]. Nanoscale, 2012, 4(10):3141-3147.
[6] 胡琼璨, 宁静恒, 刘振国, 等. 生物可用型纳米磁珠的制备和改性研究[J]. 食品与机械, 2016, 32(3):53-59.
[7] Bilal M, Zhao YP, Rasheed T, et al.Magnetic nanoparticles as versatile carriers for enzymes immobilization:A review[J]. Int J Biol Macromol, 2018, 120(Pt B):2530-2544.
[8] Cao Y, Liang XF, Li L, et al.High-efficiency EpCAM/EGFR-PLGA immunomagnetic beads with specific recognition on circulating tumor cells in colorectal cancer[J]. J Biomed Nanotechnol, 2017, 13(7):758-766.
[9] Li Y, Lim E, Field T, et al.Improving sensitivity and specificity of amyloid-β peptide and tau protein detection with anti-biofouling magnetic nanoparticles for liquid biopsy of Alzheimer’s disease
[J]. ACS Biomater Sci Eng, 2019, 5(7):3595-3605.
[10] Lin ZA, Zheng JN, Lin F, et al.Synthesis of magnetic nanoparticles with immobilized aminophenylboronic acid for selective capture of glycoproteins[J]. J Mater Chem, 2011, 21(2):518-524.
[11] Uygun DA, KaragöZler AA, AkgöL S, et al. Magnetic hydrophobic affinity nanobeads for lysozyme separation[J]. Materials Science & Engineering C, 2009, 29(7):2165-2173.
[12] Feng Z, Zhi ST, Guo L, et al.An integrated magnetic microfluidic chip for rapid immunodetection of the prostate specific antigen using immunomagnetic beads[J]. Microchim Acta, 2019, 186(4):252-265.
[13] Pan J, Yang Q.Antibody-functionalized magnetic nanoparticles for the detection of carcinoembryonic antigen using a flow-injection electrochemical device[J]. Anal Bioanal Chem, 2007, 388(1):279-286.
[14] 林梅双. 磁珠浓缩低浓度HBsAg方法的建立及应用[D]. 广州:广州医学院, 2012.
[15] 石若冰, 张志超, 陈磊, 等. 基于磁性微球的固定化金属亲和载体及其在蛋白质/多肽纯化中的应用[J]. 分析化学, 2007, 35(5):628-632.
[16] 田晶晶, 丁少华, 王红梅, 等. 纳米磁珠免疫分析技术的建立及其检测人红细胞血型Mi~a(MNS7)抗体的初步实验[J]. 中国输血杂志, 2014, 27(3):244-248.
[17] Qin SY, Yin H, Yang CL, et al.A magnetic protein biocompass[J]. Nature Materials, 2015, 15(2):217-226.
[18] Jiang M, Zhang LJ, Wang FQ, et al.Novel application of magnetic protein:Convenient one-step purification and immobilization of proteins[J]. Scientific Reports, 2017, 7(1):13329-13339.
[19] 吴明珲, 姜玲黎, 曾凡波, 等. 磁性免疫微球在人血清白蛋白纯化中的应用[J]. 药学学报, 2006, 41(7):608-614.
[20] Vertegel AA, Siegel RW, Dordick JS.Silica nanoparticle size influences the structure and enzymatic activity of adsorbed lysozyme[J]. Langmuir, 2004, 20(16):6800-6807.
[21] Norde W.Adsorption of proteins from solution at the solid-liquid interface[J]. Adv Colloid Interface Sci, 1986, 25(4):267-340.
[22] Rechendorff K, Hovgaard MB, Foss M, et al.Enhancement of protein adsorption induced by surface roughness[J]. Langmuir, 2006, 22(26):10885-10888.
[23] 李文娇, 阎俊, 杜娟, 等. 不同磁珠对过敏原蛋白纯化效果的比较研究[J]. 山东农业大学学报:自然科学版, 2017, 48(6):853-857.

六种蛋白质与生物纳米磁珠的吸附性能研究

The Study on the Adsorption Properties of Six Proteins on Biological Magnetic Nanobeads

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

[1]	李海洋, 王飞, 雷红涛, 张璇, 陈珂. 硅羟基磁珠的制备及全基因组DNA提取优化[J]. 生物技术通报, 2017, 33(6): 223-229.
[2]	任敬钢, 程超, 仲从浩, 张丽, 李荣秀. 蛋白A-葡聚糖-Fe⁴纳米磁珠的制备及对IgG分离纯化[J]. 生物技术通报, 2014, 0(7): 201-208.