NANOTECHNOLOGY-ENABLED DETECTION OF CLINICALLY RELEVANT ANTIBODIES FOR EARLY CANCER DIAGNOSIS AND IMMUNOTHERAPY MONITORING

Project Information

Nano-ImmunoEra

Grant agreement ID: 101086341

DOI

10.3030/101086341

EC signature date 17 October 2022

Start date 1 January 2023

End date 31 December 2026

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 441 600,00

Coordinated by

ALMA MATER STUDIORUM - UNIVERSITA DI BOLOGNA
Italy

Periodic Reporting for period 1 - Nano-ImmunoEra (NANOTECHNOLOGY-ENABLED DETECTION OF CLINICALLY RELEVANT ANTIBODIES FOR EARLY CANCER DIAGNOSIS AND IMMUNOTHERAPY MONITORING)

Reporting period: 2023-01-01 to 2024-12-31

Summary of the context and overall objectives of the project

Current methods for the quantitative detection of clinically-relevant antibodies mostly rely on enzyme linked immunosorbent assays (ELISAs), Western blots, and polarization assays, which are multistep, wash- and reagent-intensive processes that require sophisticated instrumentations that limit their applicability in point-of-care (POC) diagnostics and high-throughput assays. Point-of-care disposable tests instead allow more frequent diagnostic screening, especially in low in-come countries, and shortens the delay between sample acquisition, diagnosis and therapeutic decisions in clinical settings.
The goal of Nano-ImmunoEra network is to transfer the knowledge from “the bench” to real clinical practices for developing the next generation of sensing platforms able to detect multiple tumor-associated antibodies in order to improve early cancer diagnosis, effective prognosis and immunotherapy monitoring. We will demonstrate the feasibility and validate adaptability of the Nano-ImmunoEra diagnostic tools for the rapid, sensitive and multiplex detection of serological tumor-associated autoantibody and this would potentially have a significant impact on early diagnosis of cancer-affected patients at all stages. Nano-ImmunoEra platform will use as transduction mechanism, electrochemiluminescence (ECL) that -because it offers high sensitivities – is already the leading signal-transduction technique in many important clinical analyses. In ECL, the light emission is triggered by an electrochemical reaction requiring voltages lower than common batteries, without the need of lamps or lasers; for all these reasons it can allow the construction of portable, low-cost devices.
key objectives:
- Identification and optimization of selective recognition elements for clinically-relevant antibody detection; KPI1: High binding affinity (KD < 10-12 M)
- Preparation of novel nanostructured electrode material and ECL signal transduction; KPI2 costs < 1 euro/electrode.
- Understanding and optimizing of the parameters controlling the electrochemiluminescence transduction method; KPI3: “per molecule” analytical signal intensity > 100 times that of Ru(bpy)32+/TPA; Noise < 10 times less than the Ru(bpy)32+ system.
- Combining of programmable antibody-responsive nucleic acid devices and CRISPR-based amplification; KPI4: ~ 2 orders of magnitude higher sensitivity compared to direct affinity-based detection
- Real‐time monitoring of therapeutic antibodies in whole blood KPI5: Trastuzumab, Cetuximab LOQ = 10 µg ml-1; dynamic range between 5 and 180 µg ml-1
- Development of a portable ECL platform for rapid, easy to use TAA detection KPI6: anti-p53 autoantibody, anti-MUC1 autoantibody LOQ < 5 ng ml-1

The final step will be the evaluation with model samples and the comparison with the state-of-the-art biosensing technologies.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

List of the main scientific archivement of Nano-ImmunoEra
- Designed a rapid, one-pot detection of IgG antibodies (and small molecules) with high sensitivity and specificity even within complex matrices. Besides the bioanalytical applications, the switchable engineered DNA serve as programmable DNA-based tools capable of regulating Cas12a-based targeting and DNA processing in response to protein inputs and hold promise for a wide array of biotechnological applications (WP1 related achievement)
- Report the preparation of two family of electrode for electrochemiluminescence. (WP2 related achievement)
- Develop electrochemical aptasensor (EAB) for the quantification of trasduzumabin whole blood. The EAB shows a linear range within the clinical relevant concentration, fast response (<20min), potentially applicable to continuous Ab monitoring and high specificity (Kd=74 @3 7°C in whole blood) (WP3 related achievement)

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

Nano-ImmunoEra is a research and innovation staff exchange network aiming to develop innovative biosensors and diagnostic tools for detecting clinically relevant antibodies. Nano-ImmunoEra will form the next generation of clinically-oriented analytical scientists capable of delivering innovative solutions that will improve patients’ lives, reduce healthcare costs, and position Europe as a leader in biomedical devices. Since standard laboratory-based methodologies and disposable tests present several analytical limitations, we have selected antibodies as molecular targets of this project because their rapid, sensitive and specific detection in bodily fluids still represents a significant diagnostic challenge. To deliver these goals, we have assembled a highly inter- and multidisciplinary and multicultural team of European as well as strategic non-EU Countries (USA, Switzerland and South Africa) through collaboration among 7 academic institutions and 2 non-academic partners to enable ultimate impact on healthcare. The expertise of different and multidisciplinary research teams, comprised of PhD and Postdoctoral as well as experienced researchers from Universities and companies, will endow Nano-ImmunoEra with a strong intersectoral dimension, which is highly important for a broad and effective project impact. In addition, this program involves essential participation of junior researchers with exceptional scientific backgrounds and accomplishments and will provide strong support for the development of their careers. The transfer of knowledge and related training activities aim to provide scientists with the opportunity to achieve specific competencies in the field of materials sciences, nanotechnology, molecular biology and biosensors.

The future of molecular diagnostics
Our aim is to develop, optimize, and assess novel molecular diagnostic platforms with a focus on cancer monitoring. Combining different nano- and microscale materials and devices with antibody-responsive biomolecules serving as recognition elements.

The young international project
The proposed exchange program aims at boosting the academic careers of young researchers and at promoting a transfer of knowledge between Italy, USA, Spain, Germany, Switzerland, The Netherlands, and South Africa by building a multicultural team

A strong intersectoral dimension
A key aspect of Nano-ImmunoEra is its interdisciplinary nature to use a variety of approaches and strategies. The project brings together nucleic acid chemistry, molecular bioengineering, electrochemiluminescence and CRISPR-based technology.

Europe, research and careers
We aim to encourage the mobility of researchers in Europe and boost the careers of European researchers. It is essential to make Europe attractive again to the rest of the world by giving young people a taste for research and scientific careers.

International team
To deliver these goals, we have assembled a highly inter- and multidisciplinary and multicultural team of European as well as strategic non-EU Countries through collaboration among 7 academic institutions and 2 non-academic partners to enable ultimate impact on healthcare.

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