Periodic Reporting for period 1 - Nano-ImmunoEra (NANOTECHNOLOGY-ENABLED DETECTION OF CLINICALLY RELEVANT ANTIBODIES FOR EARLY CANCER DIAGNOSIS AND IMMUNOTHERAPY MONITORING)
Periodo di rendicontazione: 2023-01-01 al 2024-12-31
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.
- 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)
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.