Development Of Biosensors Research Articles

Portable Electrochemical Immunosensor Based on a Gold Microblobs-Optimized Screen-Printed Electrode for SARS-CoV-2 Diagnosis

The development of biosensors for determining the most diverse biomolecules is a constant focus of many research groups. There is a latent need to propose sensors that combine portability, simple measurements, and good analytical performance. Here, we propose an electrochemical immunosensor that is fully portable and energy-independent for diagnosing antibodies against SARS-CoV-2 (the virus that causes COVID-19). Initially, disposable screen-printed carbon electrodes (SPEs) were covered by gold microblobs (AuMBs), which were synthesized amperometrically from Au3+ ions. Then, the SPE-AuMBs were coated with cysteamine, which allowed the N-hydroxysuccinimide-activated SARS-CoV-2 antigen (spike protein) to be immobilized. The antigen-activated electrode was used to detect COVID-19 antibodies from current measurements obtained by differential pulse voltammetry. The AuMBs synthesis time was optimized, and the presence of gold structures improved the electrochemical responses of the SPE. It was possible to quantitatively determine antibodies in the concentration range of 0.25 to 10 µg mL−1. This range includes concentrations found in biological fluids from patients at any stage of the disease. An analysis took approximately the same time as traditional rapid nasal tests (20 min) and costed less, considering all the steps necessary to prepare a disposable antigen-functionalized SPE.

Responsive DNA hydrogels: design strategies and prospects for biosensing.

Hydrogels, water-filled networks that can adapt to external stimuli by altering their volume, are known for their high flexibility and biocompatibility. DNA, a critical biomolecule renowned for its exceptional characteristics including information transmission, molecular recognition, and editability, has found widespread applications in the biosensing field as well. The integration of these two biomaterials offers promising opportunities for the development of novel biosensors with enhanced sensitivity, specificity, and adaptability. Therefore, by virtue of the collective features, researchers have recently focused on the construction of responsive DNA hydrogel systems. This feature article describes recent developments in fabricating DNA hydrogels and their applications in the biosensing area. Initially, it focuses on the design strategies employed in preparing DNA hydrogels, encompassing both pure DNA hydrogels and hybridized DNA hydrogels. Subsequently, it summarizes the use of DNA hydrogels in biosensing applications, highlighting their applications in visual detection, electrochemical sensing, and optical biosensing analyses. Furthermore, the underlying responsive mechanisms within these biosensing systems are also described. Lastly, this article presents a comprehensive discussion on the existing challenges and prospects of responsive DNA hydrogels, offering insights into their potential to revolutionize the field of biosensing.

Light-triggered protease-mediated release of actin-bound cargo from synthetic cells.

Synthetic cells offer a versatile platform for addressing biomedical and environmental challenges, due to their modular design and capability to mimic cellular processes such as biosensing, intercellular communication, and metabolism. Constructing synthetic cells capable of stimuli-responsive secretion is vital for applications in targeted drug delivery and biosensor development. Previous attempts at engineering secretion for synthetic cells have been confined to non-specific cargo release via membrane pores, limiting the spatiotemporal precision and specificity necessary for selective secretion. Here, we designed and constructed a protein-based platform termed TEV Protease-mediated Releasable Actin-binding protein (TRAP) for selective, rapid, and triggerable secretion in synthetic cells. TRAP is designed to bind tightly to reconstituted actin networks and is proteolytically released from bound actin, followed by secretion via cell-penetrating peptide membrane translocation. We demonstrated TRAP's efficacy in facilitating light-activated secretion of both fluorescent and luminescent proteins. By equipping synthetic cells with a controlled secretion mechanism, TRAP paves the way for the development of stimuli-responsive biomaterials, versatile synthetic cell-based biosensing systems, and therapeutic applications through the integration of synthetic cells with living cells for targeted delivery of protein therapeutics.

Disposable glutamate biosensor based on platinum nanoparticles, carbon quantum dots and poly-L-aspartic acid

This study reports the design and development of a disposable amperometric biosensor for the determination of L-glutamate. Glutamate oxidase (GlOx) was immobilized onto a screen-printed carbon electrode (SPE) modified with poly-L-Aspartic acid (PAsp), carbon quantum dots (CQD), and platinum nanoparticles (PtNP) for the construction of the biosensor. The surface composition of the modified SPE was optimized using the one variable at a time method. The morphological properties of the biosensor were characterized by scanning electron microscopy and energy-dispersive X-ray spectroscopy. The electrochemical behavior of the modified electrodes was studied by cyclic voltammetry. Under the optimized experimental conditions the linear working range, detection limit and sensitivity of the GlOx/PtNP/CQD/PAsp/SPE were found to be 1.0 − 140 µM, 0.3 µM and 0.002 µA µM−1, respectively. The GlOx/PtNP/CQD/PAsp/SPE biosensor also exhibited good measurement repeatability. The as-developed biosensor was applied for the determination of L-glutamate in spiked serum samples and the average analytical recovery of added glutamate was 98.9 ± 3.9%.

Entropy-driven amplification reaction and the CRISPR/Cas12a system form the basis of an electrochemical biosensor for E.coli-specific detection

We present an innovative biosensor designed for the precise identification of Escherichia coli (E.coli), a predominant pathogen responsible for gastrointestinal infections. E.coli is prevalent in environments characterized by substandard water quality and can lead to severe diarrhea, especially in hospital settings. The device employs entropy-driven reactions to synthesize copious amounts of double-stranded DNA (dsDNA), which, upon binding with crRNA, triggers the CRISPR/Cas12a system’s cleavage mechanism. This process results in the separation of a ferrocene (Fc)-tagged DNA strand from the electrode, enhancing the electrochemical signal for E.coli’s rapid and accurate detection. Our tests confirm the biosensor’s ability to quantify E.coli across a dynamic range from 100 to 10 million CFU/mL, achieving a detection threshold of just over 5 CFU/mL. The development of this electrochemical biosensor highlights its exceptional selectivity, high sensitivity, and user-friendly interface for E.coli detection. It stands as a significant step forward in pathogen detection technology, promising new directions for identifying various bacterial infections through the CRISPR/Cas mechanism.

Sensing Interfaces Engineering for Organic Thin Film Transistors-Based Biosensors: Opportunities and Challenges.

Organic thin film transistors (OTFTs) enable rapid and label-free high-sensitivity detection of target analytes due to their low cost, large-area processing, biocompatibility, and inherent signal amplification. At the same time, the freedom of synthesis, tailorability, and functionalization of organic semiconductor materials and their ability to be combined with flexible substrates make them one of the ideal platforms for biosensing. However, OTFTs-based biosensors still face significant challenges, such as unexpected surface adsorption, disordered conformation, inhomogeneous graft density, and flexibility of probe molecules that biological sensing probes would face during immobilization. In this review, efficient immobilization strategies based on OTFTs biological sensing probes developed in the last 5 years are highlighted. First, the biosensors are classified according to their sensing interface. Second, a comprehensive discussion of the types of biological sensing probes is presented. Third, three commonly used methods for immobilizing biological sensing probes and their challenges are briefly described. Finally, the applications of OTFTs-based biosensors for liquid phase detection are summarized. This review provides a comprehensive and timely review of optimization in sensing interface engineering so that efficient immobilization of biological sensing probes with sensing interfaces will contribute to the development of high-performance OTFTs-based biosensors.

Overdose Detection Among High-Risk Opioid Users Via a Wearable Chest Sensor in a Supervised Injecting Facility: Protocol for an Observational Study.

Opioid overdose is a global health crisis, affecting over 27 million individuals worldwide, with more than 100,000 drug overdose deaths in the United States in 2022-2023. This protocol outlines the development of the PneumoWave chest biosensor, a wearable device being designed to detect respiratory depression in real time through chest motion measurement, intending to enhance early intervention and thereby reduce fatalities. The study aims to (1) differentiate opioid-induced respiratory depression (OIRD) from nonfatal opioid use patterns to develop and refine an overdose detection algorithm and (2) examine participants' acceptability of the chest biosensor. The study adopts an observational design over a 6-month period. The biosensor, a small device, will be worn by consenting participants during injecting events to capture chest motion data. Safe injecting facilities (SIF) in Melbourne, Victoria (site 1), and Sydney, New South Wales (site 2), which are legally sanctioned spaces where individuals can use preobtained illicit drugs under medical supervision. Each site is anticipated to recruit up to 100 participants who inject opioids and attend the SIF. Participants will wear the biosensor during supervised injecting events at both sites. The biosensor will attempt to capture data on an anticipated 40 adverse drug events. The biosensor's ability to detect OIRD will be compared to the staff-identified events that use standard protocols for managing overdoses. Measurements will include (1) chest wall movement measured by the biosensor, securely streamed to a cloud, and analyzed to refine an overdose detection algorithm and (2) acute events or potential overdose identified by site staff. Acceptability will be measured by a feedback questionnaire as many times as the participant is willing to throughout the study. As of April 2024, a total of 47 participants have been enrolled and data from 1145 injecting events have already been collected, including 10 overdose events. This consists of 17 females and 30 males with an average age of 45 years. Data analysis is ongoing. This protocol establishes a foundation for advancing wearable technology in opioid overdose prevention within SIFs. The study will provide chest wall movement data and associated overdose data that will be used to train an algorithm that allows the biosensor to detect an overdose. The study will contribute crucial insights into OIRD, emphasizing the biosensor's potential step forward in real-time intervention strategies. DERR1-10.2196/57367.

Biosensing Technologies for Detecting Legionella in Environmental Samples: A Systematic Review.

The detection of Legionella in environmental samples, such as water, is crucial for public health monitoring and outbreak prevention. Although effective, traditional detection methods, including culture-based techniques and polymerase chain reaction, have limitations such as long processing times, trained operators, and the need for specialized laboratory equipment. Biosensing technologies offer a promising alternative due to their rapid, sensitive, cost-effectiveness, and on-site detection capabilities. To summarize the current advancements in biosensor development for detecting Legionella in environmental samples, we used 'Legionella' AND 'biosensors' NEAR 'environmental samples' OR 'water' as keywords searching through the most relevant biomedical databases for research articles. After removing duplicates and inadequate articles from the n.1268 records identified using the PRISMA methodology exclusion criteria, we selected n.65 full-text articles which suited the inclusion criteria. Different results between the studies describing the current biosensing techniques, including optical, electrochemical, magnetic, and mass-sensitive sensors were observed. For each biosensing technique, sensitivity, specificity, and detection limits were evaluated. Furthermore, the integration of nanomaterials, microfluidics, and portable devices in biosensor systems' design were discussed, highlighting their role in enhancing detection performance. The potential challenges and future directions in the field of Legionella biosensing were also addressed, providing insights into the feasibility of implementing these technologies in routine environmental monitoring. Undoubtedly, biosensors can play a crucial role in the early detection and management of Legionella infections and outbreaks, ultimately protecting public health and safety.

Tetrodotoxin and the state-of-the-art progress of its associated analytical methods.

Tetrodotoxin (TTX), which is found in various marine organisms, including pufferfish, shellfish, shrimp, crab, marine gastropods, and gobies, is an effective marine toxin and the cause of many seafood poisoning incidents. Owing to its toxicity and threat to public health, the development of simple, rapid, and efficient analytical methods to detect TTX in various food matrices has garnered increasing interest worldwide. Herein, we reviewed the structure and properties, origin and sources, toxicity and poisoning, and relevant legislative measures of TTX. Additionally, we have mainly reviewed the state-of-the-art progress of analytical methods for TTX detection in the past five years, such as bioassays, immunoassays, instrumental analysis, and biosensors, and summarized their advantages and limitations. Furthermore, this review provides an in-depth discussion of the most advanced biosensors, including cell-based biosensors, immunosensors, and aptasensors. Overall, this study provides useful insights into the future development and wide application of biosensors for TTX detection.

Recent advances in portable devices for environmental monitoring applications.

Environmental pollution remains a major societal problem, leading to serious impacts on living organisms including humans. Human activities such as civilization, urbanization, and industrialization are major causes of pollution. Imposing stricter rules helps control environmental pollutant levels, creating a need for reliable pollutant monitoring in air, water, and soil. The application of traditional analytical techniques is limited in low-resource areas because they are sophisticated, expensive, and bulky. With the development of biosensors and microfluidics technology, environmental monitoring has significantly improved the analysis time, low cost, portability, and ease of use. This review discusses the fundamentals of portable devices, including microfluidics and biosensors, for environmental control. Recently, publications reviewing microfluidics and biosensor device applications have increased more than tenfold, showing the potential of emerging novel approaches for environmental monitoring. Strategies for enzyme-, immunoassay-, and molecular-based analyte sensing are discussed based on their mechanisms and applications. Microfluidic and biosensor platforms for detecting major pollutants, including metal ions, pathogens, pesticides, and antibiotic residues, are reviewed based on their working principles, advantages, and disadvantages. Challenges and future trends for the device design and fabrication process to improve performance are discussed. Miniaturization, low cost, selectivity, sensitivity, high automation, and savings in samples and reagents make the devices ideal alternatives for in-field detection, especially in low-resource areas. However, their operation with complicated environmental samples requires further research to improve the specificity and sensitivity. Although there is a wide range of devices available for environmental applications, their implementation in real-world situations is limited. This study provides insights into existing issues that can be used as references and a comparative analysis for future studies and applications.

Advances in textile-based microfluidics for biomolecule sensing.

Textile-based microfluidic biosensors represent an innovative fusion of various multidisciplinary fields, including bioelectronics, material sciences, and microfluidics. Their potential in biomedicine is significant as they leverage textiles to achieve high demands of biocompatibility with the human body and conform to the irregular surfaces of the body. In the field of microfluidics, fabric coated with hydrophobic materials serves as channels through which liquids are transferred in precise amounts to the sensing element, which in this case is a biosensor. This paper presents a condensed overview of the current developments in textile-based microfluidics and biosensors in biomedical applications over the past 20 years (2005-2024). A literature search was performed using the Scopus database. The fabrication techniques and materials used are discussed in this paper, as these will be key in various modifications and advancements in textile-based microfluidics. Furthermore, we also address the gaps in the application of textile-based microfluidic analytical devices in biomedicine and discuss the potential solutions. Advances in textile-based microfluidics are enabled by various printing and fabric manufacturing techniques, such as screen printing, embroidery, and weaving. Integration of these devices into everyday clothing holds promise for future vital sign monitoring, such as glucose, albumin, lactate, and ion levels, as well as early detection of hereditary diseases through gene detection. Although most testing currently takes place in a laboratory or controlled environment, this field is rapidly evolving and pushing the boundaries of biomedicine, improving the quality of human life.

Comparative Study of Piezoelectric MEMS for Enhanced Biosensors

Piezoelectric MEMS (Micro-Electro-Mechanical Systems) refers to devices that combine the principles of piezoelectricity with microfabrication techniques to create small-scale mechanical systems with integrated electronic components. Piezoelectric Microelectromechanical Systems (MEMS) for enhanced biosensors represent a sophisticated integration of two key technologies: piezoelectric materials and MEMS fabrication techniques. This combination results in highly sensitive, miniaturized devices that can detect and analyze biological molecules with precision. Piezoelectric MEMS devices offer unique advantages including high sensitivity, low power consumption, and miniaturization, making them ideal candidates for cutting-edge biosensor development. This article provides a comparative study of various Piezoelectric MEMS-based biosensors, highlighting their strengths and limitations.

Screen-Printed Electrodes as Low-Cost Sensors for Breast Cancer Biomarker Detection.

This review explores the emerging role of screen-printed electrodes (SPEs) in the detection of breast cancer biomarkers. We discuss the fundamental principles and fabrication techniques of SPEs, highlighting their adaptability and cost-effectiveness. The review examines various modification strategies, including nanomaterial incorporation, polymer coatings, and biomolecule immobilization, which enhance sensor performance. We analyze the application of SPEs in detecting protein, genetic, and metabolite biomarkers associated with breast cancer, presenting recent advancements and innovative approaches. The integration of SPEs with microfluidic systems and their potential in wearable devices for continuous monitoring are explored. While emphasizing the promising aspects of SPE-based biosensors, we also address current challenges in sensitivity, specificity, and real-world applicability. The review concludes by discussing future perspectives, including the potential for early screening and therapy monitoring, and the steps required for clinical implementation. This comprehensive overview aims to stimulate further research and development in SPE-based biosensors for improved breast cancer management.

The Development and Application of Biosensors in Medical Diagnostics in Indonesia

Biosensors are analytical devices that combine biological sensing elements with physicochemical detectors to provide rapid, accurate, and cost-effective solutions for medical diagnostics. Globally, these devices have revolutionized the detection and monitoring of diseases, significantly impacting patient outcomes. In Indonesia, the adoption and development of biosensor technology have accelerated over the past decade, particularly in response to the growing healthcare needs and the government's push for technological innovation. This study aims to review and analyze the development and application of biosensors in Indonesia medical diagnostics sector between 2014 and 2024. The research focuses on identifying key technological advancements, the integration of biosensors into healthcare, the challenges faced in their development and deployment, and the prospects for future growth. The study employs a comprehensive literature review and analysis of scientific publications, industry reports, and government documents related to biosensor technology in Indonesia. The review covers the evolution of biosensing materials, point-of-care testing applications, integration with digital health technologies, and regulatory frameworks. Data were synthesized to provide a detailed overview of the current state of biosensor technology and its impact on medical diagnostics in Indonesia. The findings indicate significant advancements in biosensor technology, particularly in the development of novel nanomaterials, the integration of biosensors with mobile health (mHealth) platforms, and the expansion of point-of-care testing (POCT) for infectious diseases. However, the sector faces challenges, including technological limitations, regulatory hurdles, and economic constraints, which have slowed the widespread adoption of biosensors in clinical settings. Despite these challenges, the future of biosensors in Indonesia appears promising, with continued investment and innovation expected to drive further advancements in this field.

Electronic structure engineering of N-doped carbon nanozyme via incorporating Cl and sp3-hybridized defected carbon for organophosphorus pesticides assay

Metal-free carbon-based nanozymes often exhibit superior chemical stability and detection reliability compared to their metal-doped counterparts. However, their catalytic activity remains an area ripe for further enhancement. Herein, we successfully prepared a chlorine (Cl)-modified, metal-free, and porous N-doped carbon nanozyme (Clx-pNC) via NaCl molten etching. The incorporation of Cl induced an increase in the intrinsic defects of sp3-hybridized carbon within Clx-pNC and optimized the electronic structure of the N-connected carbon atoms. Remarkably, the peroxidase (POD)-like activity of Clx-pNC was enhanced twelvefold compared to porous N-doped carbon (pNC). Theoretical simulations highlighted that the introduction of Cl not only promoted H2O2 adsorption but also lowered the energy barrier for its decomposition, facilitating the generation of active intermediates and thus boosting POD-like activity. Based on the POD mimic activity of Clx-pNC, we developed a colorimetric platform for OPs detection utilizing a cascade amplification strategy. This work provides insights into the rational design of carbon-based nanozymes and the development of nanozyme-based colorimetric biosensors.

Bio-functionalized conductive poly(acrylic acid):poly(3,4-Ethylenedioxythiophene)-Prussian blue hybrid transducer for biosensors and bioelectronics interfaces

This study presents an innovative organic-inorganic hybrid transducer of poly(acrylic acid) (PAA), poly(3,4-ethylenedioxythiophene) (PEDOT) and Prussian blue (PB) nanocatalysts. The transducer demonstrated functionality conducive to enzyme conjugation and exhibited favorable electrochemical properties for biosensor signal transduction. Fabricated by a one-tube chemical redox method, the PAA:PEDOT-PB transducer showed long-term electrocatalytic and structural stability. The performance of the transducer was characterized by high transduction activity and low charge transfer resistance, particularly for H2O2 reduction. It achieves a linear detection range from 1.0 μM to 4.0 mM, with a sensitivity of 494 ± 10 μA mM−1 cm−1 and a LOD of 0.34 μM. The PAA:PEDOT-PB transducer featured a high density of carboxyl groups (DCOOH = 14.64 ± 0.05 μmol cm−2) that promoted the immobilization of the H2O2-dependent oxidase enzyme lactate oxidase (LOx) with an EDC/S–NHS coupling agent. The LOx-PAA:PEDOT-PB transducer was developed for lactate biosensing. The transducer provided high LOx-enzyme affinity (KMapp = 1.47 ± 0.05 mM), and a rapid response time (10 s) for lactate detection across a concentration range of 5.0 μM to 4.0 mM, showing a sensitivity of 223 ± 3 μA mM−1 cm−1 and an LOD of 1.45 μM. The LOx-PAA:PEDOT-PB transducer was integrated with a flexible screen-printed electrode, incorporating a wireless, battery-free near-field communication potentiostat module to measure lactate in artificial sweat on a skin model via smartphone. The PAA:PEDOT-PB transducer could enable connections with multiple bio-recognition molecules through polycarboxylic acid groups, providing potential avenues for the development of advanced biosensors.

Using biosensors for the early diagnosis of arboviruses: a scoping review

Mapping scientific knowledge about the development of biosensors to be used for the early diagnosis of arboviruses. Scoping review based on studies searched in Lilacs, PubMed, SciELO and Web of Science databases. Nineteen (19) articles were selected, in total. The selected studies presented different types of biosensors used to detect arboviruses and their variants, with emphasis on dengue and Zika viruses. Electrochemical, optical and plasmon resonance were the biosensors mostly used for the aforementioned purpose due to their little margin of error. They enable reliable and fast diagnosis processes, besides being easy to handle and its low cost. Biosensors capable of identifying arboviruses are not yet available in health services, since they still require advancements to be achieved at practical research stages.

Development of an electrochemical biosensor for non-invasive cholesterol monitoring via microneedle-based interstitial fluid extraction

In this study, we present the development of an innovative electrochemical biosensor integrated into a microneedle-based system for non-invasive and sensitive quantification of cholesterol levels in interstitial fluid (ISF). The biosensor employs a graphene-based electrode with a polyelectrolyte interlayer to immobilize cholesterol oxidase (ChOx), enabling selective cholesterol detection. Graphene oxide is electrochemically reduced to form a conductive layer, and PANI is chosen as the optimal polyelectrolyte for ChOx immobilization. The biosensor's performance is thoroughly evaluated, demonstrating excellent sensitivity, stability, and selectivity. Furthermore, the biosensor is successfully applied to skin-mimicking agarose gel and porcine skin, showcasing its potential for real-world interstitial fluid extraction and cholesterol monitoring. The integrated microneedle-based system offers a promising approach for non-invasive monitoring of cholesterol levels, with implications for personalized healthcare diagnostics.

Multi-walled carbon nanotubes functionalized with a new Schiff base containing phenylboronic acid residues: applicationtothe development of a bienzymatic glucose biosensor using a response surface methodology approach.

Aninnovative supramolecular architectureis reportedfor bienzymatic glucose biosensing based on the use of a nanohybrid made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with a Schiff base modified with two phenylboronic acid residues (SB-dBA) as platform for the site-specific immobilization of the glycoproteins glucose oxidase (GOx) and horseradish peroxidase (HRP). The analytical signal was obtained from amperometric experiments at - 0.050V in the presence of 5.0 × 10-4M hydroquinone as redox mediator. The concentration of GOx and HRP and the interaction time between the enzymes and the nanohybrid MWCNT-SB-dBA deposited at glassy carbon electrodes (GCEs) were optimized through a central composite design (CCD)/response surface methodology (RSM). The optimal concentrations of GOx and HRP were 3.0mgmL-1 and 1.50mgmL-1, respectively, while the optimum interaction time was 3.0min. The bienzymatic biosensor presented a sensitivity of (24 ± 2) × 102 µA dL mg-1 ((44 ± 4) × 102 µA M-1), a linear range between 0.06mg dL-1 and 21.6mg dL-1 (3.1µM-1.2mM) (R2 = 0.9991),and detection and quantification limits of 0.02mg dL-1 (1.0µM) and 0.06mg dL-1 (3.1µM), respectively. The reproducibility for five sensors prepared with the same MWCNT-SB-dBA nanohybrid was 6.3%, while the reproducibility for sensors prepared with five different nanohybrids and five electrodes each was 7.9%. The GCE/MWCNT-SB-dBA/GOx-HRP was successfully used for the quantification of glucose in artificial human urine and commercial human serum samples.

Enhanced pyrophosphate detection: Utilizing oPD-derived carbon dots and Fe3+ interactions in a paper strip biosensor

The development of portable, cost-effective, and straightforward DNA biosensors holds immense importance in various fields, including healthcare, environmental monitoring, and food safety. This study contributes to the objective by introducing an innovative approach for synthesizing carbon dots (Cdots) with high quantum yield (QY) and remarkable selectivity for Fe3+ ions. Utilizing o-phenylenediamine as a precursor, the study achieved a straightforward and environmentally friendly synthesis method, enabling the efficient detachment of metal ions from the Cdot surface upon introducing pyrophosphate (PPi). The presence of surface hydroxyl and amino groups facilitated specific Fe3+ recognition. Employing D-optimal response surface methodology, the study optimized Cdot synthesis parameters, identifying temperature and heating time as critical factors influencing QY. Statistical analysis confirmed the model's reliability, predicting maximum QY of 48.8 % with minimal deviation from experimental results. Characterization studies revealed the amorphous nature of Cdots through HR-TEM, XRD, and FTIR analysis. Furthermore, the proposed LAMP/PPi biosensing technique demonstrated higher sensitivity, specificity, and repeatability, with negligible interference from common anions and efficacy across varying pH levels. The limit of detection (LOD) of 0.079 (±0.01) μM and the detection range of 0.1 μM–2 mM underscore the biosensor's practical utility. This study highlights a promising direction for developing paper-based LAMP/PPi biosensors with potential diagnostics and environmental monitoring applications. Significantly, the biosensing technique is applicable to any DNA amplification method generating pyrophosphate (PPi) as a by-product.