Citation: HE Xiao-Qin, LUO Li, GUO Lei, XIE Jian-Wei. An Aptameric Biolayer Interferometric Assay for Detection of Recombinant Human Erythropoietin-α[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(5): 670-675. doi: 10.19756/j.issn.0253-3820.191676 shu

An Aptameric Biolayer Interferometric Assay for Detection of Recombinant Human Erythropoietin-α

HE Xiao-Qin ^1,2, ,
LUO Li ^1,3 ,
GUO Lei ¹ ,
XIE Jian-Wei ¹

1.
State Key Laboratory of Toxicology and Medical Countermeasures, Institute of Toxicology and Pharmacology, Academy of Military Medical Sciences, Beijing 100850, China;
2.
Department of Spine Surgery and Institute for Orthopaedic Research, the 2^nd Clinical Medical College of Jinan University and the 1^st Affiliated Hospital of Southern University of Science and Technology(Shenzhen People's Hospital), Shenzhen 518020, China;
3.
School of Public Health, Hebei Medical University, Shijiazhuang 050017, China

Received Date: 15 November 2019
Revised Date: 24 March 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21175152 and 21974152).

The biolayer interferometry (BLI), one promising biosensing technique for real-time, high-sensitive detection of analytes, is a new surface-sensitive spectrometric technique in which the change of biolayer thickness is reflected by measuring the interferometric phase deflections. In our previous research, we selected an ssDNA aptamer, 39 nt, with definite interaction characteristics for recombinant human erythropoietin-α (EPO-α). In this work, a new aptameric BLI assay of EPO-α based on this aptamer 39 nt was established. Firstly, the biotinylated aptamer 39 nt was immobilized onto the optical fiber streptavidin biosensor tip, and the analytes in microliter volume were quickly diffused via high-speed oscillation at 2200 r/min. Then, the key parameters such as the orientation of aptamer sequence, the concentration for immobilization, and the overcome of nonspecific adsorption were investigated and optimized to exhibit the affinity recognition of aptamers, and it was found that the 3’-end biotinylated aptamer 39 nt with 50 nmol/L provided the best sensitivity, and the addition of Tween 20 and bovine serum albumin (BSA) could efficiently avoid nonspecific adsorption effect. Besides, towards the original designed disposable biosensor tip, the affinity between EPO-α and aptamer 39 nt for ca. 10 times was maintained by optimizing the regeneration condition. Furthermore, a signal amplification approach of a sandwiched format for detection of EPO-αvia wheat germ agglutinin (WGA) was established, with the linear range of 10-200 nmol/L, and a limit of detection of 5 nmol/L. This assay was further applied to the measurement of EPO-α in 50% human plasma with recoveries of 86.7%-104.2% and RSD less than 11%, providing a satisfied result. The BLI sensing system developed here provides a helpful way for practical applications for label-free protein interaction and detection.
- Aptamer,
- Recombinant human erythropoietin-α,
- Biolayer interferometry,
- Wheat germ agglutinin,
- Signal amplification
1. [1]
2. [2]
3. [3]
4. [4]
5. [5]
6. [6]
7. [7]
8. [8]
9. [9]
10. [10]
11. [11]
12. [12]
13. [13]
14. [14]
15. [15]
1. [1]
  Qilong Fang , Yiqi Li , Jiangyihui Sheng , Quan Yuan , Jie Tan . Magical Pesticide Residue Detection Test Strips: Aptamer-based Lateral Flow Test Strips for Organophosphorus Pesticide Detection. University Chemistry, 2024, 39(5): 80-89. doi: 10.3866/PKU.DXHX202310004
2. [2]
  Shuyu Liu , Xiaomin Sun , Bohan Song , Gaofeng Zeng , Bingbing Du , Chongshen Guo , Cong Wang , Lei Wang . Design and Fabrication of Phospholipid-Vesicle-based Artificial Cells towards Biomedical Applications. University Chemistry, 2024, 39(11): 182-188. doi: 10.12461/PKU.DXHX202404113
3. [3]
  Yixuan Zhu , Qingtong Wang , Jin Li , Lin Chen , Junlong Zhao . Blog of Oxytocin. University Chemistry, 2024, 39(9): 134-140. doi: 10.12461/PKU.DXHX202310090
4. [4]
  Zhaoxin LI , Ruibo WEI , Min ZHANG , Zefeng WANG , Jing ZHENG , Jianbo LIU . Advancements in the construction of inorganic protocells and their cell mimic and bio-catalytical applications. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2286-2302. doi: 10.11862/CJIC.20240235
5. [5]
  Yukun Xing , Xiaoyu Xie , Fangfang Chen . A Sunlit Gift: Vitamin D. University Chemistry, 2024, 39(9): 28-34. doi: 10.12461/PKU.DXHX202402006
6. [6]
  Jijun Sun , Qianlang Wang , Qian Chen , Quanqin Zhao , Shumei Zhai . The Antibiotic Legion’s Manifesto to Human Allies. University Chemistry, 2025, 40(4): 307-321. doi: 10.12461/PKU.DXHX202405206
7. [7]
  Yikai Wang , Xiaolin Jiang , Haoming Song , Nan Wei , Yifan Wang , Xinjun Xu , Cuihong Li , Hao Lu , Yahui Liu , Zhishan Bo . 氰基修饰的苝二酰亚胺衍生物作为膜厚不敏感型阴极界面材料用于高效有机太阳能电池. Acta Physico-Chimica Sinica, 2025, 41(3): 2406007-. doi: 10.3866/PKU.WHXB202406007
8. [8]
  Qilu DU , Li ZHAO , Peng NIE , Bo XU . Synthesis and characterization of osmium-germyl complexes stabilized by triphenyl ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1088-1094. doi: 10.11862/CJIC.20240006
9. [9]
  Zhongxin YU , Wei SONG , Yang LIU , Yuxue DING , Fanhao MENG , Shuju WANG , Lixin YOU . Fluorescence sensing on chlortetracycline of a Zn-coordination polymer based on mixed ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2415-2421. doi: 10.11862/CJIC.20240304
10. [10]
  Linjie ZHU , Xufeng LIU . Electrocatalytic hydrogen evolution performance of tetra-iron complexes with bridging diphosphine ligands. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 321-328. doi: 10.11862/CJIC.20240207
11. [11]
  Zijuan LI , Xuan LÜ , Jiaojiao CHEN , Haiyang ZHAO , Shuo SUN , Zhiwu ZHANG , Jianlong ZHANG , Yanling MA , Jie LI , Zixian FENG , Jiahui LIU . Synthesis of visual fluorescence emission CdSe nanocrystals based on ligand regulation. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 308-320. doi: 10.11862/CJIC.20240138
12. [12]
  Yaling Chen . Basic Theory and Competitive Exam Analysis of Dynamic Isotope Effect. University Chemistry, 2024, 39(8): 403-410. doi: 10.3866/PKU.DXHX202311093
13. [13]
  Xiao SANG , Qi LIU , Jianping LANG . Synthesis, structure, and fluorescence properties of Zn(Ⅱ) coordination polymers containing tetra-alkenylpyridine ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2124-2132. doi: 10.11862/CJIC.20240158
14. [14]
  Hongjie SHEN , Haozhe MIAO , Yuhe YANG , Yinghua LI , Deguang HUANG , Xiaofeng ZHANG . Synthesis, crystal structure, and fluorescence properties of two Cu(Ⅰ) complexes based on pyridyl ligand. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 855-863. doi: 10.11862/CJIC.20250009
15. [15]
  Xiaoning TANG , Junnan LIU , Xingfu YANG , Jie LEI , Qiuyang LUO , Shu XIA , An XUE . Effect of sodium alginate-sodium carboxymethylcellulose gel layer on the stability of Zn anodes. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1452-1460. doi: 10.11862/CJIC.20240191
16. [16]
  Zhenhua Wang , Haoyang Feng , Xiaoyang Shao , Wenru Fan . Vitamins in Solid Propellants: Controlled Synthesis of Neutral Macromolecular Bonding Agents. University Chemistry, 2025, 40(4): 1-9. doi: 10.3866/PKU.DXHX202401007
17. [17]
  Cun WANG , Shaohan XU , Yuqian ZHANG , Yaoyao ZHANG , Tao GONG , Rong WEN , Yuhang LIAO , Yanrong REN . Terbium complex electrochemiluminescent emitters: Synthesis and application in the detection of epinephrine. Chinese Journal of Inorganic Chemistry, 2025, 41(7): 1351-1360. doi: 10.11862/CJIC.20240427
18. [18]
  Xiaoling LUO , Pintian ZOU , Xiaoyan WANG , Zheng LIU , Xiangfei KONG , Qun TANG , Sheng WANG . Synthesis, crystal structures, and properties of lanthanide metal-organic frameworks based on 2, 5-dibromoterephthalic acid ligand. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1143-1150. doi: 10.11862/CJIC.20230271
19. [19]
  Yingchun ZHANG , Yiwei SHI , Ruijie YANG , Xin WANG , Zhiguo SONG , Min WANG . Dual ligands manganese complexes based on benzene sulfonic acid and 2, 2′-bipyridine: Structure and catalytic properties and mechanism in Mannich reaction. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1501-1510. doi: 10.11862/CJIC.20240078
20. [20]
  Youlin SI , Shuquan SUN , Junsong YANG , Zijun BIE , Yan CHEN , Li LUO . Synthesis and adsorption properties of Zn(Ⅱ) metal-organic framework based on 3, 3', 5, 5'-tetraimidazolyl biphenyl ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1755-1762. doi: 10.11862/CJIC.20240061

Get Citation

PDF

XML

Metrics

PDF Downloads(8)
Abstract views(646)
HTML views(18)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142