The present fellowship proposes the development and characterization of electrochemical platforms to monitor the quantitative detection of clinically-relevant antibodies, that will be rapid, specific, convenient and selective enough to be employed directly in complex samples, such as blood serum and whole blood.
The detection of specific antibodies and other diagnostic proteins plays a crucial role in the diagnosis of many diseases, infections and pathologies and despite their widespread use current detection methods are either cumbersome, multi-step and laboratory-bound processes (i.e. ELISA, western blot) or only qualitative or semi-quantitative (i.e. later flow immunoassay). Due to the above considerations, better analytical tools that allow the rapid, inexpensive, and quantitative measurement of clinically-relevant biomarkers, including antibodies, are urgently needed. To address this need, during my Marie-Skłodowska Curie project (“DNA-NANO-AB”), I have developed different DNA-based sensors and devices for the detection of a wide range of molecules, including antibodies, that may be of utility for diagnostic applications.
To do so, I employ the designability and versatility of synthetic DNA sequences those can be used as scaffolds to create antibody-responsive nanodevices and I take advantage of the positive feature of electrochemical detection that include interference-free measurements in complex matrices, low-cost instrumentation and mass-producible sensors.
To reach the proposed objectives I will undertake the following tasks:
1. Rational design and characterization of different DNA-based strategies (conformational change, proximity-based approach, etc.) to improve target measurement efficiency using spectroscopic and electrochemical-based approaches. Different DNA-based elements will be tested as well as several recognition element/antibody couples.
2. Development and optimization of versatile electrochemical platforms for the orthogonal and multiplexed antibodies detection. Systematic study of: i) redox reporters, ii) antigen tags, iii) electrochemical parameters, iv) signal amplification.
3. Development of platforms using disposable screen-printed electrodes produced in house to reduce the fabrication cost of each sensor.
4. Test of representative real samples using the developed electrochemical platform. The platforms will be firstly challenged with blood serum and ultimately with whole blood.
In conclusion “DNA-NANO-AB” has paved the basis for new classes of DNA-based sensors for antibody detection. Specifically, the possibility of coupling the advantages of synthetic nucleic acids with the positive feature of electrochemical detection and the possible amplification processes to enhance the sensibility of the sensors make the sensing platforms developed during this project well suited for point-of-care applications.
