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Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition
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Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition

A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensor and Bioelectronic Devices".

Deadline for manuscript submissions: closed (20 April 2025) | Viewed by 17292

Special Issue Editor

Dr. Zhuangqiang Gao

E-Mail Website
Guest Editor
Marshall Laboratory of Biomedical Engineering, Research Center for Biosensor and Nanotheranostic, School of Biomedical Engineering, Health Science Center, Shenzhen University, Shenzhen 518060, China
Interests: chemical and biological sensors; optical sensors; metal nanomaterials; catalysis; plasmonics; microfluidics; disease diagnosis; environmental monitoring
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

After the success of the first Volume of the Special Issue entitled “Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis”, the Journal’s Editorial Board, in collaboration with the Guest Editor, Dr. Zhuangqiang Gao, is launching Volume II under the same title. This Special Issue continues to explore the exciting developments and applications of nano biosensors in in vivo/vitro diagnosis and any other area of biological interest.

Over the past two years, there has been a notable trend toward the integration of novel nanostructures with innovative sensing principles to enhance the analytical performance of biosensors. Researchers have focused on designing and fabricating advanced nanostructures with unique properties, such as electrical, magnetic, optical, and plasmonic activities, to enable the highly sensitive and selective detection of biomarkers.

Volume II encompasses a diverse range of topics, including the synthesis and characterization of novel nanostructures, the development of innovative biosensors with high analytical performance, the exploration of new sensing principles, the identification of promising diagnostic biomarkers, and the emerging applications of nano biosensors in both in vivo and in vitro diagnosis. Through the contributions of researchers in the field, this volume aims to advance our understanding of nano biosensors and their potential impact on healthcare and disease diagnosis.

Dr. Zhuangqiang Gao
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biosensors is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2200 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • nanostructures
  • nanomaterials
  • nanodevices
  • biosensors
  • biomarkers
  • in vivo/vitro diagnosis

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Published Papers (10 papers)

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Research

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18 pages, 4658 KiB  
Article
Atomic Pt-Layer-Coated Au Peroxidase Nanozymes with Enhanced Activity for Ultrasensitive Colorimetric Immunoassay of Interleukin-12
by Han Zhang, Xiang Peng, Hao Song, Yongfeng Tan, Jianglian Xu, Qunfang Li and Zhuangqiang Gao
Biosensors 2025, 15(4), 239; https://doi.org/10.3390/bios15040239 - 9 Apr 2025
Viewed by 306
Abstract
Interleukin-12 (IL-12), a crucial biomarker for immune and inflammatory responses, plays a pivotal role in diagnosing and managing diverse pathological conditions. Although colorimetric enzyme-linked immunosorbent assays (CELISAs) have been extensively employed to detect IL-12 in biological samples, their sensitivity is inherently limited by [...] Read more.
Interleukin-12 (IL-12), a crucial biomarker for immune and inflammatory responses, plays a pivotal role in diagnosing and managing diverse pathological conditions. Although colorimetric enzyme-linked immunosorbent assays (CELISAs) have been extensively employed to detect IL-12 in biological samples, their sensitivity is inherently limited by the catalytic efficiency of enzyme labels, presenting substantial challenges in achieving ultrasensitive detection and enabling pre-symptomatic diagnosis of diseases. In this study, we address this limitation by developing a novel peroxidase nanozyme, featuring ultrathin Pt skins consisting of only ~4 atomic layers, coated on Au nanoparticles (denoted as Au@Pt4LNPs). These Au@Pt4LNPs exhibit remarkable catalytic performance, achieving a ~1063-fold enhancement in peroxidase-like activity compared to horseradish peroxidase (HRP), while minimizing Pt consumption, thereby improving Pt utilization efficiency and reducing costs. This advancement facilitates the construction of an ultrasensitive CELISA capable of detecting IL-12 at femtomolar concentrations. Using Au@Pt4LNPs as the signal labels, the developed CELISA demonstrates a quantitative detection range from 0.1 to 100 pg mL−1, with a limit of detection (LOD) as low as 0.084 pg mL−1 (1.1 fM), offering ~10 times greater sensitivity than the HRP-based CELISA. This study highlights the potential of Au@Pt4LNP nanozymes as advanced signal labels, opening new avenues for next-generation ultrasensitive bioassays. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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19 pages, 5726 KiB  
Article
MOX Nanosensors to Detect Colorectal Cancer Relapses from Patient’s Blood at Three Years Follow-Up, and Gender Correlation
by Michele Astolfi, Giulia Zonta, Cesare Malagù, Gabriele Anania and Giorgio Rispoli
Biosensors 2025, 15(1), 56; https://doi.org/10.3390/bios15010056 - 16 Jan 2025
Viewed by 1037
Abstract
Colorectal cancer represents 10% of all the annual tumors diagnosed worldwide, being often not timely diagnosed, because its symptoms are typically lacking or very mild. Therefore, it is crucial to develop and validate innovative low-invasive techniques to detect it before becoming intractable. To [...] Read more.
Colorectal cancer represents 10% of all the annual tumors diagnosed worldwide, being often not timely diagnosed, because its symptoms are typically lacking or very mild. Therefore, it is crucial to develop and validate innovative low-invasive techniques to detect it before becoming intractable. To this aim, a device equipped with nanostructured gas sensors has been employed to detect the airborne molecules of blood samples collected from healthy subjects, and from colorectal cancer affected patients at different stages of their pre- and post-surgery therapeutic path. Data was scrutinized by using statistical standard techniques to highlight their statistical differences, and through principal component analysis and support vector machine to classify them. The device was able to readily distinguish between the pre-surgery blood samples (i.e., taken when the patient had cancer), and the ones up to three years post-surgery (i.e., following the tumor removal) or the ones from healthy subjects. Finally, the correlation of the sensor responses with the patient/healthy subject’s gender was investigated, resulting negligible. These results pave the path toward a clinical validation of this device to monitor the patient’s health status by detecting possible relapses, to parallel to clinical follow-up protocols. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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13 pages, 3759 KiB  
Article
An MSRE-Assisted Glycerol-Enhanced RPA-CRISPR/Cas12a Method for Methylation Detection
by Zhiquan Lu, Zilu Ye, Ping Li, Yike Jiang, Sanyang Han and Lan Ma
Biosensors 2024, 14(12), 608; https://doi.org/10.3390/bios14120608 - 12 Dec 2024
Cited by 1 | Viewed by 1166
Abstract
Background: Nasopharyngeal carcinoma (NPC) is a malignant tumor with high prevalence in southern China. Aberrant DNA methylation, as a hallmark of cancer, is extensively present in NPC, the detection of which facilitates early diagnosis and prognostic improvement of NPC. Conventional methylation detection methods [...] Read more.
Background: Nasopharyngeal carcinoma (NPC) is a malignant tumor with high prevalence in southern China. Aberrant DNA methylation, as a hallmark of cancer, is extensively present in NPC, the detection of which facilitates early diagnosis and prognostic improvement of NPC. Conventional methylation detection methods relying on bisulfite conversion have limitations such as time-consuming, complex processes and sample degradation; thus, a more rapid and efficient method is needed. Methods: We propose a novel DNA methylation assay based on methylation-sensitive restriction endonuclease (MSRE) HhaI digestion and Glycerol-enhanced recombinase polymerase amplification (RPA)-CRISPR/Cas12a detection (HGRC). MSRE has a fast digestion rate, and HhaI specifically cleaves unmethylated DNA at a specific locus, leaving the methylated target intact to trigger the downstream RPA-Cas12a detection step, generating a fluorescence signal. Moreover, the detection step was supplemented with glycerol for the separation of Cas12a-containing components and RPA- and template-containing components, which avoids over-consumption of the template and, thus, enhances the amplification efficiency and detection sensitivity. Results: The HGRC method exhibits excellent performance in the detection of a CNE2-specific methylation locus with a (limit of detection) LOD of 100 aM and a linear range of 100 aM to 100 fM. It also responds well to different methylation levels and is capable of distinguishing methylation levels as low as 0.1%. Moreover, this method can distinguish NPC cells from normal cells by detecting methylation in cellular genomes. This method provides a rapid and sensitive approach for NPC detection and also holds good application prospects for other cancers and diseases featuring DNA methylation as a biomarker. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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15 pages, 7613 KiB  
Article
Electrochemical Analysis of Amyloid Plaques and ApoE4 with Chitosan-Coated Gold Nanostars for Alzheimer’s Detection
by Min-Kyung Shin, Ariadna Schuck, Minhee Kang and Yong-Sang Kim
Biosensors 2024, 14(10), 510; https://doi.org/10.3390/bios14100510 - 17 Oct 2024
Cited by 2 | Viewed by 1697
Abstract
Monitoring the progression of Alzheimer’s disease (AD) is crucial for mitigating dementia symptoms, alleviating pain, and improving mobility. Traditionally, AD biomarkers like amyloid plaques are predominantly identified in cerebrospinal fluid (CSF) due to their concentrated presence. However, detecting these markers in blood is [...] Read more.
Monitoring the progression of Alzheimer’s disease (AD) is crucial for mitigating dementia symptoms, alleviating pain, and improving mobility. Traditionally, AD biomarkers like amyloid plaques are predominantly identified in cerebrospinal fluid (CSF) due to their concentrated presence. However, detecting these markers in blood is hindered by the blood–brain barrier (BBB), resulting in lower concentrations. To address this challenge and identify pertinent AD biomarkers—specifically amyloid plaques and apolipoprotein E4 (ApoE4)—in blood plasma, we propose an innovative approach. This involves enhancing a screen-printed carbon electrode (SPCE) with an immobilization matrix comprising gold nanostars (AuNSs) coated with chitosan. Morphological and electrical analyses confirmed superior dispersion and conductivity with 0.5% chitosan, supported by UV–Vis spectroscopy, cyclic voltammetry, and Nyquist plots. Subsequent clinical assays measured electrical responses to quantify amyloid-β 42 (Aβ42) (15.63–1000 pg/mL) and APoE4 levels (0.41 to 40 ng/mL) in human blood plasma samples. Differential pulse voltammetry (DPV) responses exhibited peak currents proportional to biomarker concentrations, demonstrating high linear correlations (0.985 for Aβ42 and 0.919 for APoE4) with minimal error bars. Cross-reactivity tests with mixed solutions of amyloid-β 40 (Aβ40), Aβ42, and ApoE4 indicated minimal interference between biomarkers (<3% variation), further confirming the high specificity of the developed sensor. Validation studies demonstrated a strong concurrence with the gold-standard enzyme-linked immunosorbent assay (ELISA), while interference tests indicated a minimal variation in peak currents. This improved device presents promising potential as a point-of-care system, offering a less invasive, cost-effective, and simplified approach to detecting and tracking the progression of AD. The substantial surface binding area further supports the efficacy of our method, offering a promising avenue for advancing AD diagnostics. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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11 pages, 5333 KiB  
Article
Fluorescence Immunoassay of Prostate-Specific Antigen Using 3D Paddle Screw-Type Devices and Their Rotating System
by Su Bin Han, Han Sol Kim, Young Ju Jo and Soo Suk Lee
Biosensors 2024, 14(10), 494; https://doi.org/10.3390/bios14100494 - 11 Oct 2024
Viewed by 1370
Abstract
In this paper, we present a sensitive and highly reproducible fluorescence immunosensor for detecting PSA in human serum. A unique feature of this study is that it uses creatively designed paddle screw-type devices and their custom-made rotating system for PSA immunoassay. The paddle [...] Read more.
In this paper, we present a sensitive and highly reproducible fluorescence immunosensor for detecting PSA in human serum. A unique feature of this study is that it uses creatively designed paddle screw-type devices and their custom-made rotating system for PSA immunoassay. The paddle screw devices were designed to maximize the surface-to-volume ratio over which the immunoassay reaction could occur to improve detection sensitivity. This paddle screw-based immunoassay offers an accessible and efficient method with a short analysis time of less than 30 min. Active rotation of the paddle screw plays a crucial role in fast and accurate analysis of PSA. Additionally, a paddle screw-based immunoassay and subsequent fluorescence detection using a custom prototype fluorescence detection system were compared to a typical well plate-based immunoassay system. Results of PSA detection in human serum showed that the detection sensitivity through the paddle screw-based analysis improved about five times compared to that with a well plate-based analysis. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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13 pages, 3048 KiB  
Article
Paper-Based DNA Biosensor for Rapid and Selective Detection of miR-21
by Alexander Hunt, Sri Ramulu Torati and Gymama Slaughter
Biosensors 2024, 14(10), 485; https://doi.org/10.3390/bios14100485 - 8 Oct 2024
Cited by 2 | Viewed by 2329
Abstract
Cancer is the second leading cause of death globally, with 9.7 million fatalities in 2022. While routine screenings are vital for early detection, healthcare disparities persist, highlighting the need for equitable solutions. Recent advancements in cancer biomarker identification, particularly microRNAs (miRs), have improved [...] Read more.
Cancer is the second leading cause of death globally, with 9.7 million fatalities in 2022. While routine screenings are vital for early detection, healthcare disparities persist, highlighting the need for equitable solutions. Recent advancements in cancer biomarker identification, particularly microRNAs (miRs), have improved early detection. MiR-21 is notably overexpressed in various cancers and can be a valuable diagnostic tool. Traditional detection methods, though accurate, are costly and complex, limiting their use in resource-limited settings. Paper-based electrochemical biosensors offer a promising alternative, providing cost-effective, sensitive, and rapid diagnostics suitable for point-of-care use. This study introduces an innovative electrochemical paper-based biosensor that leverages gold inkjet printing for the quantitative detection of miR-21. The biosensor, aimed at developing cost-effective point-of-care devices for low-resource settings, uses thiolated self-assembled monolayers to immobilize single-stranded DNA-21 (ssDNA-21) on electrodeposited gold nanoparticles (AuNPs) on the printed gold surface, facilitating specific miR-21 capture. The hybridization of ssDNA-21 with miR-21 increases the anionic barrier density, impeding electron transfer from the redox probe and resulting in a current suppression that correlates with miR-21 concentration. The biosensor exhibited a linear detection range from 1 fM to 1 nM miR-21 with a sensitivity of 7.69 fM µA−1 cm2 and a rapid response time (15 min). With a low detection limit of 0.35 fM miR-21 in serum, the biosensor also demonstrates excellent selectivity against interferent species. This study introduces an electrochemical paper-based biosensor that uses gold inkjet printing to precisely detect miR-21, a key biomarker overexpressed in various cancers. This innovative device highlights the potential for cost-effective, accessible cancer diagnostics in underserved areas. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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Review

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19 pages, 885 KiB  
Review
Piezoelectric Chemosensors and Biosensors in Medical Diagnostics
by Miroslav Pohanka
Biosensors 2025, 15(3), 197; https://doi.org/10.3390/bios15030197 - 20 Mar 2025
Viewed by 1431
Abstract
This article explores the development and application of innovative piezoelectric sensors in point-of-care diagnostics. It highlights the significance of bedside tests, such as lateral flow and electrochemical tests, in providing rapid and accurate results directly at the patient’s location. This paper delves into [...] Read more.
This article explores the development and application of innovative piezoelectric sensors in point-of-care diagnostics. It highlights the significance of bedside tests, such as lateral flow and electrochemical tests, in providing rapid and accurate results directly at the patient’s location. This paper delves into the principles of piezoelectric assays, emphasizing their ability to detect disease-related biomarkers through mechanical stress-induced electrical signals. Various applications of piezoelectric chemosensors and biosensors are discussed, including their use in the detection of cancer biomarkers, pathogens, and other health-related analytes. This article also addresses the integration of piezoelectric materials with advanced sensing technologies to improve diagnostic accuracy and efficiency, offering a comprehensive overview of current advances and future directions in medical diagnostics. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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23 pages, 3427 KiB  
Review
Development of Neurodegenerative Disease Diagnosis and Monitoring from Traditional to Digital Biomarkers
by Jaeyoon Song, Eunseo Cho, Huiseop Lee, Suyoung Lee, Sehyeon Kim and Jinsik Kim
Biosensors 2025, 15(2), 102; https://doi.org/10.3390/bios15020102 - 11 Feb 2025
Cited by 1 | Viewed by 1380
Abstract
Monitoring and assessing the progression of symptoms in neurodegenerative diseases, including Alzheimer’s and Parkinson’s disease, are critical for improving patient outcomes. Traditional biomarkers, such as cerebrospinal fluid analysis and brain imaging, are widely used to investigate the underlying mechanisms of disease and enable [...] Read more.
Monitoring and assessing the progression of symptoms in neurodegenerative diseases, including Alzheimer’s and Parkinson’s disease, are critical for improving patient outcomes. Traditional biomarkers, such as cerebrospinal fluid analysis and brain imaging, are widely used to investigate the underlying mechanisms of disease and enable early diagnosis. In contrast, digital biomarkers derived from phenotypic changes—such as EEG, eye movement, gait, and speech analysis—offer a noninvasive and accessible alternative. Leveraging portable and widely available devices, such as smartphones and wearable sensors, digital biomarkers are emerging as a promising tool for ND diagnosis and monitoring. This review highlights the comprehensive developments in digital biomarkers, emphasizing their unique advantages and integration potential alongside traditional biomarkers. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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14 pages, 1032 KiB  
Review
Needle-Shaped Biosensors for Precision Diagnoses: From Benchtop Development to In Vitro and In Vivo Applications
by Ruier Xue, Fei Deng, Tianruo Guo, Alexander Epps, Nigel H. Lovell and Mohit N. Shivdasani
Biosensors 2024, 14(8), 391; https://doi.org/10.3390/bios14080391 - 13 Aug 2024
Viewed by 2298
Abstract
To achieve the accurate recognition of biomarkers or pathological characteristics within tissues or cells, in situ detection using biosensor technology offers crucial insights into the nature, stage, and progression of diseases, paving the way for enhanced precision in diagnostic approaches and treatment strategies. [...] Read more.
To achieve the accurate recognition of biomarkers or pathological characteristics within tissues or cells, in situ detection using biosensor technology offers crucial insights into the nature, stage, and progression of diseases, paving the way for enhanced precision in diagnostic approaches and treatment strategies. The implementation of needle-shaped biosensors (N-biosensors) presents a highly promising method for conducting in situ measurements of clinical biomarkers in various organs, such as in the brain or spinal cord. Previous studies have highlighted the excellent performance of different N-biosensor designs in detecting biomarkers from clinical samples in vitro. Recent preclinical in vivo studies have also shown significant progress in the clinical translation of N-biosensor technology for in situ biomarker detection, enabling highly accurate diagnoses for cancer, diabetes, and infectious diseases. This article begins with an overview of current state-of-the-art benchtop N-biosensor designs, discusses their preclinical applications for sensitive diagnoses, and concludes by exploring the challenges and potential avenues for next-generation N-biosensor technology. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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19 pages, 5361 KiB  
Review
Aptamer Screening: Current Methods and Future Trend towards Non-SELEX Approach
by Zhihui Fang, Xiaorui Feng, Fan Tang, Han Jiang, Shuyuan Han, Ran Tao and Chenze Lu
Biosensors 2024, 14(7), 350; https://doi.org/10.3390/bios14070350 - 18 Jul 2024
Cited by 6 | Viewed by 3570
Abstract
Aptamers are nucleic acid sequences that specifically bind with target molecules and are vital to applications such as biosensing, drug development, disease diagnostics, etc. The traditional selection procedure of aptamers is based on the Systematic Evolution of Ligands by an Exponential Enrichment (SELEX) [...] Read more.
Aptamers are nucleic acid sequences that specifically bind with target molecules and are vital to applications such as biosensing, drug development, disease diagnostics, etc. The traditional selection procedure of aptamers is based on the Systematic Evolution of Ligands by an Exponential Enrichment (SELEX) process, which relies on repeating cycles of screening and amplification. With the rapid development of aptamer applications, RNA and XNA aptamers draw more attention than before. But their selection is troublesome due to the necessary reverse transcription and transcription process (RNA) or low efficiency and accuracy of enzymes for amplification (XNA). In light of this, we review the recent advances in aptamer selection methods and give an outlook on future development in a non-SELEX approach, which simplifies the procedure and reduces the experimental costs. We first provide an overview of the traditional SELEX methods mostly designed for screening DNA aptamers to introduce the common tools and methods. Then a section on the current screening methods for RNA and XNA is prepared to demonstrate the efforts put into screening these aptamers and the current difficulties. We further predict that the future trend of aptamer selection lies in non-SELEX methods that do not require nucleic acid amplification. We divide non-SELEX methods into an immobilized format and non-immobilized format and discuss how high-resolution partitioning methods could facilitate the further improvement of selection efficiency and accuracy. Full article
(This article belongs to the Special Issue Nano Biosensors and Their Applications for In Vivo/Vitro Diagnosis—2nd Edition)
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