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引用本文:   杨歌, 韩诗邈, 赵丽萍, 朱超, 黄渊余, 屈锋. 毛细管电泳法筛选人IgG的Fc片段核酸适配体. 分析化学, 2020, 48(5): 601-607. doi:  10.19756/j.issn.0253-3820.191761 [复制]

Citation:   YANG Ge , HAN Shi-Miao , ZHAO Li-Ping , ZHU Chao , HUANG Yuan-Yu , QU Feng . Screening of Aptamer for Human IgG Fc Fragment by Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment. Chinese Journal of Analytical Chemistry, 2020, 48(5): 601-607. doi: 10.19756/j.issn.0253-3820.191761 [复制]

毛细管电泳法筛选人IgG的Fc片段核酸适配体

收稿日期: 2019-12-24

基金项目: 本文系国家自然科学基金项目(Nos.21675012,21874010,21827810)资助

Screening of Aptamer for Human IgG Fc Fragment by Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment

Received Date:  2019-12-24

Fund Project:  This work was supported by the National Natural Science Foundation of China (Nos. 21675012, 21874010, 21827810).

基于毛细管电泳-指数富集配体系统进化技术(CE-SELEX)建立了免疫球蛋白G(IgG)的Fc片段的核酸适配体筛选方法。通过毛细管区带电泳方法分析IgG完整蛋白、Fc与Fab片段的样品的纯度、荷电性质及是否吸附的特征,并比较三者与核酸库的亲和力。结果表明,IgG与核酸库形成明显的复合物,而Fc和Fab片段与核酸库的亲和力很弱。在筛选Fc片段的适配体时,选择Fab片段作为反筛靶,对去除非特异性结合序列无显著作用。设计了针对Fc片段的适配体筛选方案,在优化的孵育条件(10 mmol/L Na2HPO4-KH2PO4 (pH 7.17),0.05 mmol/L Mg2+,37℃孵育25 min)下,经3轮筛选获得Fc片段的核酸适配体Seq Fc1-3,KD值为0.071-0.321 μmol/L,经胶体金比色分析方法验证了其亲和力与特异性。

关键词:   免疫球蛋白, 免疫球蛋白G Fc片段, 核酸适配体筛选, 毛细管电泳, 毛细管电泳-指数富集配体系统进化
Key words:   Immunoglobulin, IgG Fc fragment, Aptamer screening, Capillary electrophoresis, Capillary electrophoresis-systematic evolution of ligands by exponential enrichment
[1]

Chan K R, Ong E Z, Mok D Z, Ooi E E. Expert. Rev. Anti-Infect. Ther.,2015,13(11): 1351-1360

[2]

Roopenian D C, Akilesh S. Nat. Rev. Immunol.,2007,7(9): 715-725

[3]

Fonseca M H G, Furtado G P, Bezerra M R L, Pontes L Q, Fernandes C F C. Int. J. Biol. Macromol., 2018,119: 306-311

[4]

Shen Y, Li H, Zhao L, Li G, Chen B, Guo Q, Gao B, Wu J, Yang T, Jin L, Su Y. PLoS One, 2017,12(8): e0183326

[5]

Suzuki T, Ishii-Watabe A, Tada M, Kobayashi T, Kanayasu-Toyoda T, Kawanishi T, Yamaguchi T. J. Immunol.,2010,184(4): 1968-1976

[6]

Shan L, Colazet M, Rosenthal K L, Yu X Q, Bee J S, Ferguson A, Damschroder M M, Wu H, Dall’Acqua W F, Tsui P, Oganesyan V. PLoS One,2016,11 (8): e0160345

[7]

Quast I, Peschke B, Lünemann Jan D. Cell. Mol. Life Sci.,2017,74(5):837-847

[8]

Li L, Tong R, Chu H, Wang W, Langer R, Kohane D S. Proc. Natl. Acad. Sci. USA,2014,111(48): 17099-17103

[9]

Gordon C K L, Eisenstein M, Soh H T. ACS Sens.,2018,(3): 2574-2580

[10]

Zumrut H E, Ara M N, Fraile M, Maio G, Mallikaratchy P. Nucleic Acid Ther.,2016,26 (3): 190-198

[11]

Moccia F, Platella C, Musumeci D, Batool S, Zumrut H, Bradshaw J, Mallikaratchy P, Montesarchio D. Int. J. Biol. Macromol.,2019,133: 839-849

[12]

Mendonsa S D, Bowser M T. Anal. Chem.,2004,76(18): 5387-5392

[13]

Miyakawa S, Nomura Y, Sakamoto T, Yamaguchi Y, Kato K, Yamazaki S, Nakamura Y. RNA,2008,14(6): 1154-1163

[14]

Berezovski M, Drabovich A, Krylova S M, Musheev M, Okhonin V, Petrov A, Krylov S N. J. Am. Chem. Soc.,2005,127(9): 3165-3171

[15]

Berezovski M V, Musheev M U, Drabovich A P, Jitkova J V, Krylov S N. Nat. Protoc.,2006,1(3): 1359-1369

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毛细管电泳法筛选人IgG的Fc片段核酸适配体

杨歌, 韩诗邈, 赵丽萍, 朱超, 黄渊余, 屈锋

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