Jožef Stefan Institute · Public Research Call 2025

NANO-SENSE

Nanoengineered Graphitic Carbon Nitride Electrochemical Biosensors for Advanced Biomolecular Interactions, detecting pesticide residues at femtomolar to picomolar sensitivity.

36 moDuration

fM-pMDetection Range

5 WPsWork Packages

4 TeamsCollaborating

150kEUR/yrBudget

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01 · Overview

Project Abstract

NANO-SENSE develops a next-generation electrochemical biosensor platform based on nanoengineered graphitic carbon nitride nanosheets, engineered via cold plasma technology, for ultrasensitive detection of chlorpyrifos and related pesticide residues in food and environmental matrices.

Graphitic carbon nitride is a promising alternative to conventional 2D nanomaterials such as graphene, MXene, and carbon nanotubes. With a high bandgap energy of 2.76 eV, inherent porosity, abundant active sites, metal-free composition, and eco-friendly synthesis, g-CN represents an ideal transducer material.

Using cold plasma surface engineering, the project generates targeted functional groups on few-layer g-CN nanosheets without compromising structural integrity. These groups enable covalent, oriented antibody immobilization via EDC/NHS chemistry, creating a highly selective immunosensor with detection limits in the femtomolar to picomolar range.

The combination of plasma-modified g-CN surface chemistry and antibody-based detection creates a highly efficient, cost-effective, and sustainable sensing platform validated against ELISA and GC-MS in real fruit juice matrices.

Key Technologies

Target Analyte

Validation Methods

Sample Matrices

Project Novelty

No prior literature combines g-CN with plasma-engineered modifications for biosensing. NANO-SENSE fills this gap with a greener, scalable, and reproducible platform.

TRL Progression

TRL 2-3
WP1-2

TRL 3-4
WP3

TRL 4-5
WP4

02 · Research Goals

Specific Objectives

SO1

⬡

g-CN Synthesis

Develop a facile protocol for the synthesis of g-CN nanosheets with selective properties using triazine, amide and urea-based monomers. Optimize reaction conditions, temperatures, and durations to tailor structural and functional characteristics.

SO2

⚡

Plasma Tailoring

Systematic optimization of plasma parameters: gas composition, pressure (0.1-10 Torr), power (10-500 W), and treatment duration (1-30 min) for controlled surface modification and functional group generation on g-CN.

SO3

◈

Hybrid Composites

Strategically integrate metal nanoparticles, transition metal sulphides or oxides, along with compatible polymers, onto the g-CN surface to optimize synergistic properties for multifunctional sensing applications.

SO4

⊡

Electrode Fabrication

Fabricate screen-printed working electrodes with functionalized g-CN nanosheets and optimize controlled immobilization of anti-chlorpyrifos antibodies via EDC/NHS chemistry.

SO5

◎

Ultrasensitive Detection

Validate NANO-SENSE specificity and sensitivity in complex real-world matrices and achieve reliable, rapid detection of chlorpyrifos at trace levels with cross-validation against ELISA and GC-MS.

03 · Methodology

Work Packages

WP 1

Synthesis of Graphitic Carbon Nitride

Month 1 -> 30 · TRL 3-4

JSI-O2 · NIC

T 1.1

Thermal and chemical assisted synthesis with calcination, ball-milling, and thermal oxidation to produce few-layer nanosheets using Melamine, Thiourea, and Dicyandiamide precursors.

T 1.2

Plasma-assisted production using microwave and radiofrequency plasma systems with parametric optimization of pressure, power, gas flow, and substrate temperature.

T 1.3

Physicochemical characterization through SEM, AFM, TEM, XPS, Raman, THz, FTIR, XRD, and water contact angle measurements.

WP 2

Surface Engineering and Hybrid Design of g-CN

Month 8 -> 24 · TRL 2-3

JSI-F6

T 2.1

Plasma-enabled surface engineering with heteroatom doping using N2, O2, NH3, Ar, S, H2, and CO2 to target more than 10 at% functional group concentration.

T 2.2

Chemical functionalization via conducting polymer grafting and diazonium chemistry for covalent attachment of multifunctional molecules.

T 2.3

Hybrid g-CN and metallic composites with transition metal sulphides and oxides, including crystal phase optimization via dry plasma processing.

WP 3

Electrode Functionalization and Biomolecular Interactions

Month 13 -> 30 · TRL 2-3

JSI-O2 · JSI-K7

T 3.1

Functionalization of working electrodes through g-CN electrodeposition on commercial SPEs and CV activation with optimized deposition conditions.

T 3.2

Biomolecular interaction and parametric optimization with EDC/NHS bioconjugation, Anti-CP antibody immobilization, and BSA blocking.

T 3.3

Morphology and electrochemical characterization at every functionalization step using SEM, contact angle, CV, and DPV.

WP 4

Validation and Calibration of Biosensor Platform

Month 18 -> 36 · TRL 4-5

JSI-O2 · JSI-K7

T 4.1

Electrochemical analysis using CV, DPV, and chronoamperometry with fabrication parameter optimization across antibody concentration, scan rate, pH, and temperature.

T 4.2

Real sample testing in spiked orange, apple, and pomegranate juice, including sample preparation and interference studies with related pesticides.

T 4.3

Validation and calibration via LOD/LOQ determination, repeatability, precision, stability assessment, and cross-validation with ELISA and GC-MS.

WP 5

Management, Dissemination and Communications

Month 1 -> 36 · Cross-cutting

JSI-O2

T 5.1

Project and risk management including PI oversight, consortium agreement, progress meetings, mitigation monitoring, and financial reporting.

T 5.2

Exploitation and IP with six research articles, two short communications, five international conferences, and a potential patent on plasma surface enhancement.

T 5.3

Communication and public engagement through the project website, newsletters, open days, researcher outreach, and young researcher involvement.

04 · Research Team

Task Leaders & Collaborators

Principal Investigator

Dr. Raghuraj Singh Chouhan

PI · WP1, WP3, WP4 Lead

Dept. of Environmental Sciences (JSI-O2)

Jožef Stefan Institute

NanomaterialsBiosensorsBioconjugationPesticide Detectionh-index 22

Prof. Dr. Uroš Cvelbar

WP2 Lead

Dept. of Gaseous Electronics (JSI-F6)

Jožef Stefan Institute

Plasma NanoscienceMaterials ScienceEnergy Storage

NMS

Dr. Neelakandan M. Santhosh

Post-Doctoral Researcher · WP2

Dept. of Gaseous Electronics (JSI-F6)

Jožef Stefan Institute

Low-Pressure PlasmaCarbon NanostructuresElectrochemistry

KŽ

Dr. Kristina Žagar Soderžnik

Task Leader · WP3, WP4

Dept. of Nanostructured Materials (JSI-K7)

Jožef Stefan Institute

Electrochemical SensorsBiosensorsToxic Organic Compounds

Dr. Jelena Vujancevic

Post-Doctoral Researcher · WP3, WP4

Dept. of Nanostructured Materials (JSI-K7)

Jožef Stefan Institute

Thin-Film FabricationData AnalysisProcess Optimization

Prof. Dr. Ester Heath

Collaborator · WP5 Validation

Group of Organic Analysis (JSI-O2)

Jožef Stefan Institute

GC-MS/MSLC-MS/MSTrace Organics

Dr. David Heath

Collaborator · WP5 Validation

Dept. of Environmental Sciences (JSI-O2)

Jožef Stefan Institute

Analytical ChemistryPesticide Residue Analysis30+ Years Experience

Assis. Prof. Dr. Ivan Jerman

Collaborator · WP1 Characterization

National Institute of Chemistry (NIC)

Ljubljana

NanoparticlesNano-CoatingsRaman / AFM / SEM

Prof. Dr. Milena Horvat

Advisory & WP5 Management

Dean, JSI Postgraduate School

Head, Dept. Environmental Sciences

Trace-Level DetectionProject Management30+ Years Experience

07 · Performance Metrics

Key Performance Indicators

KPI	Metric	Target Result
Control Synthesis of g-CN	Few-layered nanosheets thickness and yield	2-4 nm thickness · 90-95% yield
Speed of Plasma Tailoring System	Produced functionalized g-CN per minute	200-250 mg/min
Concentration of Functional Groups	Atomic percentage on g-CN surface	> 10 at%
Electrochemical Biosensor Platform	Integrated g-CN with biological components	fM to pM/mL detection range
Data Analysis and Publication	High-impact peer-reviewed output	>= IF 10 · 6 articles + 2 short comms
Conference Dissemination	International presentations	5 conferences (E-MRS, ECS, Biosensors...)
IP and Patents	Plasma surface enhancement technique	1 patent application (projected)