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Colorimetric determination of the early biomarker hypoxia-inducible factor-1 alpha (HIF-1α) in circulating exosomes by using a gold seed-coated with aptamer-functionalized Au@Au core-shell peroxidase mimic

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Abstract

An ultra-sensitive method is described here for the determination of HIF-1α (an early biomarker for myocardial infarction) in circulating exosomes in serum. Gold nanospheres were functionalized with a HIF-1α-binding aptamer via sulfydryl chemistry. The apt-AuNP-coated gold seeds were grown by seed-mediated growth, and this significantly increased the peroxidase-mimicking property the nanoparticles. A chromogenic system composed of 3,3′5,5′-tetramethylbenzidine and hydrogen peroxide was used. Absorbance at 652 nm increases linearly in the 0.3 to 200 ng L−1 HIF-1α concentration range, and the limit of detection is 0.2 ng L−1. The method was tested by analyzing rat serum from isoproterenol (ISO)-induced myocardial infarction. It allows HIF-1α to be directly determined in a 25 μL sample without preconcentration. The assay is not interfered by the polydispersity of exosomes released under either health and disease conditions.

Gold nanospheres were functionalized with a HIF-1α-binding aptamer via sulfydryl chemistry. Nanosized gold seed particles were then modified with the functionalized gold nanospheres, and this strongly increases the peroxidase-mimicking activity of the nanomaterial. By using the tetramethylbenzidine/H2O2 chromogenic system, the absorbance at 652 nm increases linearly in the 0.3 to 200 ng L-1 HIF-1α concentration range.

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Abbreviations

CVDs:

Cardiovascular diseases

MI:

Myocardial infarction

cTn:

Cardiac troponin

LDH:

Lactate dehydrogenase

CK-MB:

Creatine kinase MB

HIF-1α:

Hypoxia-inducible factor-1 alpha

HRP:

Horseradish peroxidase

ISEV:

International Society for Extracellular Vesicles

AuNPs:

Gold nanoparticles

LOD:

Limit of detection

NTA:

Nanoparticle tracking analysis

DLS:

Dynamic light scattering

TEM:

Transmission electron microscope

CV:

Coefficient of variation

ISO:

Isoproterenol hydrochloride

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Acknowledgements

CAS “Light of West China” Program (No. 2018XBZG_XBQNXZ_A_005), Biological Resources Program, Chinese Academy of Sciences (KFJ-BRP-008)

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Authors and Affiliations

  1. State Key Laboratory for Quality Research in Chinese Medicines, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China

    Qian-Long Wang, Li Chen, Chon-Kit Lio, Hua Zhou & Pei Luo

  2. Centre for Natural Products Research, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, China

    Qian-Long Wang & Lin-Sen Qing

  3. State Key Laboratory of Bioorganic and Natural Products Chemistry, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China

    Wei-Xue Huang & Pu-Juan Zhang

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  1. Qian-Long Wang
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  2. Wei-Xue Huang
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  4. Li Chen
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Correspondence to Lin-Sen Qing or Pei Luo.

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Wang, QL., Huang, WX., Zhang, PJ. et al. Colorimetric determination of the early biomarker hypoxia-inducible factor-1 alpha (HIF-1α) in circulating exosomes by using a gold seed-coated with aptamer-functionalized Au@Au core-shell peroxidase mimic. Microchim Acta 187, 61 (2020). https://doi.org/10.1007/s00604-019-4035-z

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