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Citation: LUO Yu, WANG Teng-Fei, LI Wen-Jing, WANG Jin-E, PEI Ren-Jun. Selection and Characterization of Single Stranded DNA Aptamer for Human Coagulation Factor 7a[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(7): 1068-1074. doi: 10.19756/j.issn.0253-3820.191204 shu

Selection and Characterization of Single Stranded DNA Aptamer for Human Coagulation Factor 7a

LUO Yu ^1,2, ,
WANG Teng-Fei ¹ ,
LI Wen-Jing ^1,2 ,
WANG Jin-E ¹ ,
PEI Ren-Jun ¹

1.
Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou 215123, China;
2.
Nano Science and Technology Institute, University of Science and Technology of China, Suzhou 215123, China

Received Date: 15 April 2019
Revised Date: 5 May 2019

Fund Project: This work was supported by the Novo Nordisk-CAS Research Fund (No.NNCAS-2014-8), the National Natural Science Foundation of China(No.21775160)and the Science Foundation of Jiangsu Province (No.BE2016680).

Human coagulation factor 7a (FVⅡa) is the core factor for exogenous coagulation pathway. Developing safe, effective and economical coagulants or anticoagulants is still a challenge. In this study, a single-stranded DNA (ssDNA) aptamer for FVⅡa was isolated based on systematic evolution of ligands by exponential enrichment (SELEX), which was based on the isolation of the sequences bound to the target protein, which would be absorbed on nitrocellulose membrane. The aptamer was isolated after 11 rounds of selection. The apparent dissociation constant of the aptamer to FVⅡa was 102 nmol/L. The aptamer-based fluorescent assay for FVⅡa was established, exhibing simplicity and efficiency. The quantitative detection range of FVⅡa by this assay was 10-30 nmol/L, providing a new strategy for detection of FVⅡa. The non-denaturation polyacrylamide gel electrophoresis and the aptamer-based fluorescent assay results both showed that the aptamer had good selectivity toward FVⅡa.
- Coagulation factor 7a,
- Aptamer,
- Systematic evolution of ligands by exponential enrichment,
- Selection,
- Fluorescent assay
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