Determination of cell metabolite VEGF₁₆₅ and dynamic analysis of protein-DNA interactions by combination of microfluidic technique and luminescent switch-on probe

Xuexia Lin; Ka-Ho Leung; Ling Lin; Luyao Lin; Sheng Lin; Chung-Hang Leung; Dik-Lung Ma; Jin-Ming Lin

doi:10.1016/j.bios.2015.11.089

Determination of cell metabolite VEGF₁₆₅ and dynamic analysis of protein-DNA interactions by combination of microfluidic technique and luminescent switch-on probe

Biosens Bioelectron. 2016 May 15:79:41-7. doi: 10.1016/j.bios.2015.11.089. Epub 2015 Nov 30.

Authors

Xuexia Lin¹, Ka-Ho Leung², Ling Lin³, Luyao Lin³, Sheng Lin², Chung-Hang Leung⁴, Dik-Lung Ma⁵, Jin-Ming Lin⁶

Affiliations

¹ Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China; College of Chemical Engineering, Huaqiao University, Xiamen 361000, China; Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.
² Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.
³ Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China.
⁴ State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.
⁵ Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China. Electronic address: edmondma@hkbu.edu.hk.
⁶ Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China. Electronic address: jmlin@mail.tsinghua.edu.cn.

PMID: 26686922
DOI: 10.1016/j.bios.2015.11.089

Abstract

In this paper, we rationally design a novel G-quadruplex-selective luminescent iridium (III) complex for rapid detection of oligonucleotide and VEGF165 in microfluidics. This new probe is applied as a convenient biosensor for label-free quantitative analysis of VEGF165 protein from cell metabolism, as well as for studying the kinetics of the aptamer-protein interaction combination with a microfluidic platform. As a result, we have successfully established a quantitative analysis of VEGF165 from cell metabolism. Furthermore, based on the principles of hydrodynamic focusing and diffusive mixing, different transient states during kinetics process were monitored and recorded. Thus, the combination of microfluidic technique and G-quadruplex luminescent probe will be potentially applied in the studies of intramolecular interactions and molecule recognition in the future.

Keywords: Functional nucleic acid; Iridium (III) probe; Kinetic analysis; Microfluidic device; VEGF(165).

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Aptamers, Nucleotide / chemistry*
Biosensing Techniques / methods
Cell Line
Coordination Complexes / chemistry*
G-Quadruplexes
Humans
Iridium / chemistry*
Limit of Detection
Luminescence
Luminescent Agents / chemistry*
Luminescent Measurements / methods
Microfluidic Analytical Techniques / methods
Vascular Endothelial Growth Factor A / analysis*

Substances

Aptamers, Nucleotide
Coordination Complexes
Luminescent Agents
VEGFA protein, human
Vascular Endothelial Growth Factor A
Iridium