Citation: YANG Ge, ZHAO Yi, HAN Shi-Miao, ZHU Chao, HUANG Yuan-Yu, QU Feng. Screening Aptamer of Apo-transferrin via Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment and Environmental Factors Analysis[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(5): 632-641. doi: 10.19756/j.issn.0253-3820.191694 shu

Screening Aptamer of Apo-transferrin via Capillary Electrophoresis-Systematic Evolution of Ligands by Exponential Enrichment and Environmental Factors Analysis

YANG Ge ^1,2, ,
ZHAO Yi ¹ ,
HAN Shi-Miao ¹ ,
ZHU Chao ¹ ,
HUANG Yuan-Yu ^1,2 ,
QU Feng ¹

1.
School of Life Science, Beijing Institute of Technology, Key Laboratory of Molecular Medicine and Biological Diagnosis, Ministry of Industry and Information Technology, Beijing 100081, China;
2.
Advanced Research Institute of Multidisciplinary Science, Beijing Institute of Technology, Beijing 100081, China

Received Date: 25 November 2019
Revised Date: 12 December 2019

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

Aptamer is oligonucleotide with high affinity and specificity toward the target obtained via systematic evolution of ligands by exponential enrichment (SELEX). However, due to the complexity of environmental factors and the unknown influence on the selection and application of aptamers, the optimization of multiple environmental factors through conventional experiments requires a huge amount of work and samples, so there are few systematic studies. Capillary electrophoresis (CE)-SELEX has become a highly efficient technology for aptamers screening because of the advantages of CE in microscale, high efficiency and low cost. In this study, Human apo-transferrin (A-TF) was used as a model to study the effects of factors based on CE, including length of ssDNA library, incubation temperature, type and pH of incubation buffer, metal ions etc. The results showed that the shorter ssDNA library had higher affinity with the target protein, the incubation temperature, type and pH of incubation buffer affected the formation of complex, and the low concentrations of K^＋, Ca²⁺ and Mg²⁺ metal ions promoted the formation of complexes. The A-TF aptamer Seq A3 obtained by three rounds of selection had a dissociation constant (K_D) of 0.476 μmol/L, and its affinity and specificity were verified by CE-laser induced fluorescence in human serum environment. The results indicated that Seq A3 could specifically recognize A-TF in serum.
- Apo-transferrin,
- Aptamer screening,
- Influencing factors,
- Capillary electrophoresis,
- Systematic evolution of ligands by exponential enrichment
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]
16. [16]
17. [17]
18. [18]
19. [19]
20. [20]
21. [21]
22. [22]
23. [23]
24. [24]
25. [25]
26. [26]
27. [27]
28. [28]
29. [29]
30. [30]
31. [31]
32. [32]
33. [33]
34. [34]
35. [35]
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