A sensitive biosensor based on a ferrocene-marked adapter for the fluorescence detection of platelet-derived growth factor BB

https://doi.org/10.1016/j.jlumin.2020.117042Get rights and content

Highlights

  • Sensitive and simple β-cyclodextrin-modified carbon dot fluorescent nanoscale biosensors were synthesized.

  • A fluorescent nanoprobe system integrating specific host–guest identification and photoinduced electron transfer was created.

  • Ferrocene-Marked Adapter with double properties of adapter and electron receptor was developed.

  • The adapter served as a sensitive and selective biological receptor for detecting platelet-derived growth factor BB.

Abstract

A novel fluorescent nanometer biosensor integrating a β-cyclodextrin-modified carbon dot (β-CD-CD) fluorescent nanoprobe and ferrocene-marked adapter (ssDNA-Fc) was developed and used to measure platelet-derived growth factor (PDGF)-BB. When no PDGF-BB was added, ssDNA-Fc induced the enhanced fluorescence of β-CD-CD nanoprobes. After adding PDGF-BB, ssDNA-Fc specifically bonded with PDGF-BB. Meanwhile, the β-CD-CDs and Fc in ssDNA-Fc initiated host–guest identification, which then induced photoinduced electron transfer and decreased fluorescence, thereby contributing to the high-sensitivity monitoring of PDGF-BB. This novel approach showed a wide linear range of 10 pg mL−1 to 8 μg mL−1 and a low detection limit of 6 pg mL−1 (RSD = ±4.6%). Moreover, the spiked recovery rate in human serum was 98.2%–106.2% (RSD ≤ ±3.79%), indicating that this method can be used to monitor PDGF-BB in human serum.

Graphical abstract

Scheme 1. Schematic of β-CD-CD fluorescent nanoprobe for PDGF-BB detection.

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Introduction

Platelet-derived growth factor (PDGF)-BB is a well-known serum cytokine that regulates cell growth, division, and transformation [1,2]. PDGF-BB is a critical cancer-related protein marker in the human body with direct and indirect participation in cell proliferation, cell transformation, and tumor growth[3,4]. PDGF-BB is also a good predictor and indicator of early renal function deterioration, including diabetes, prostate cancer, and breast cancer[5,6]. Precise qualitative and quantitative detection of PDGF-BB can help in the early diagnosis and treatment of relevant diseases [7], as well as contribute to bioresearch and clinical diagnosis. Currently, various approaches such as enzyme-linked immunosorbent assay [8], antibody-based radioisotopes [9], molecular fluorescence technique [10], high-sensitivity electrochemical process [11], colorimetry [12], Raman method [13], and chemiluminescence immunoassay [14] are used to measure PDGF-BB. Despite the high sensitivity of these methods, they exhibit several limitations, such as high cost, instrumental complexity, and high requirement for operators. Thus, designing and creating convenient, sensitive, and selective biosensors with fast response and low cost are of great importance.

Carbon quantum dots (CDs) [15] are novel nanomaterials attracting extensive interest because of their advantages, such as high water solubility, low toxicity, high biological compatibility, light stability, abundant electron–hole exciton pairs, and functionality [16,17]. These CDs are favorable electron donors or receptors with strong ability of photoinduced electron transfer (PET)[18], synthesis and marked ability [19], and optical properties [20]. Moreover, CDs have wide-ranging applications, such as biological imaging [21], food safety [22], biomedicine[23], and chemical sensing. However, CDs from direct preparation usually have insufficient fluorescent quantum yield (FLQY < 50%) and sensitivity. For this reason, we doped N atoms containing quinquevalent electrons into the CDs[24,25], forming N-doped CDs to improve the FLQY[26]. β-Cyclodextrin (β-CD) is a ring-shaped oligonucleotide characterized by unique hydrophobic cavity, hydrophilic external structure, high water solubility [27], biocompatibility, and low toxicity [28]. The hydrophobic cavity of β-CD enables organic substances, inorganic materials, and biological molecules with appropriate sizes to be selectively coated through physical and chemical forces [29], thereby forming stable host–guest inclusion complexes, such as ferrocene[30] and adapters [31]. Accordingly, β-CD is attracting considerable attention, but its chemical inertia complicates the qualitative or quantitative analysis of substances that enter the cavity of β-CD. Nevertheless, β-CD functionalized CDs that integrate the properties of supramolecular compounds and nanometer properties of CDs can be prepared [30,32]. Molecular recognition and qualitative or quantitative detection can be achieved through PET or fluorescence resonance energy transfer of CDs.

Adapters are a type of functional nucleic acid macromolecules with specificity and affinity equivalent to antigen antibodies and can bind with various target substances, such as organic micromolecules [33], metal ions [34], proteins [35], and living cells [36]. Adapters with low cost, high stability, and high target-identification ability can be easily synthesized, marked, and modified in recent years; thus, they are extensively used in biomedicine, such as in biosensor development and nanoparticle-based drug delivery[37]. Ferrocene (Fc) is a common substance [38] that can penetrate the holes of β-CD-CDs through hydrophobic interaction, forming a stable 1:1 complex compound [39] and thus shortening the distance from the CDs of the probes [30]. When the distance between Fc and CDs is sufficiently close, under fluorescent irradiation, the excited electrons of CDs migrate to the empty orbit of Fc and do not return to the ground state, thereby inducing the PET and weakening the fluorescence of CDs [30,40]. When Fc is marked onto an adapter (e.g., single-stranded DNA (ssDNA)), the double properties of these substances can be obtained, including the specific molecular-recognition ability of the adapter and the electron signal transduction ability of Fc [41,42]. Both substances can interact with the holes of β-CD through host–guest identification, forming inclusions [31,39,43].

ssDNA and Fc can compete for host–guest in the cavity of β-CD, and ssDNA can block the PET between Fc and CDs. On the basis of this phenomenon and the double properties of ssDNA-Fc, we designed a novel fluorescent nano-bioaptamer sensor for the highly sensitive and rapid detection of PDGF-BB.

Section snippets

Materials and instruments

PDGF-BB was purchased from Cyagen Biosciences, Suzhou, China. PDGF-AA and PDGF-AB were purchased from Beijing Biolab Technology Co., Ltd. Mono(6-amino-6-deoxy)-β-CD was purchased from Shandong Binzhou Zhiyuan Biotechnology Co., Ltd. Anhydrous citric acid (CA), ethylenediamine, phosphate buffer solution (PBS; pH 7.4; containing 137 mM NaCl; 2.68 mM KCl, 8.1 mM Na2HPO4, and 1.76 mM KH2PO4), (ferrocenylmethyl)trimethylammonium iodide (Fc), and bovine serum albumin (BSA) were provided by Shanghai

Characterization

The β-CD-CD fluorescent nanoprobes were characterized by FTIR, UV–vis spectrum, TEM, and XPS.

The FTIR spectrum of β-CD-CDs exhibited three intense peaks at 1156, 1083, and 1031 cm1 (Fig. 1A), which were characteristic of β-CD and caused by the coupling extensional vibration of C–O–H, C–C, and C–O and the asymmetric glucosidic bond vibration of C–O–C. The peak at 1717 cm1 almost disappeared in CDs due to the stretching vibration of –COOH. However, two new peaks at 1684 and 1554 cm1 appearing

Conclusions

Novel β-CD-CDs were synthesized starting from β-CD-functionalized environment-friendly N-doped CDs as fluorescent nanomaterials and used to measure PDGF-BB on the basis of the specific binding between the adapter and PDGF-BB and the photoelectron migration between cyclodextrin cavity and ferrocene. The PDGF-BB detection range widened according to the adapter-induced enhancement of fluorescence in β-CD-CDs, showing a linear range of 10 pg mL−1–8 μg mL−1 and a low detection limit of 6 pg mL−1

Declaration of competing interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed influencing the position presented in, or the review of, the manuscript entitled.

Acknowledgment

This work was financially supported by the Graduate Scientific Research Foundation of Shanxi Normal University (0109–01053002).

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