Label-free, real-time detection platform of Hepatitis B Virus antigens with protein biosensors
Chronic Hepatitis B virus (HBV) is the cause for severe liver pathologies leading to ~ 1 million deaths per year, and Romania has among the highest incidence among EU countries. Screening and early clinic diagnosis are extremely relevant to prevent further viral spreading. Importantly, early productive infection can be evidenced by measuring the titers of the secretory HBV proteins, the “surface” (HBs) or “early” (HBe) antigens (Ag). The HBsAg is the first serological antigen detectable in serum ~ one week after HBV infection, followed by HBeAg, which becomes evident during viral replication. This proposal aims to place innovation at the heart of discovery, and develop a portable, label-free, first-generation protein-based nanosensor, for the sensitive and selective detection of HBeAg, by exploiting its interaction with the specific anti-HBe antibody. At the core of the endeavor lies an innovative sensing paradigm, according to which analytes of interest are identified as they disrupt the ionic current passing through a single nanopore. Knowing that nanopore technology evolved into a mature, single-molecule screening technology and revolutionized single-molecule detection, as done in Europe by Oxford Nanopore Technologies (https://nanoporetech.com), our platform will pave the way to developing nearly ideal sensors for HVB detection, for integration in low-cost, low-powered, portable, and wearable devices. Our project will start at technological readiness level, as we have preliminary data demonstrating the feasibility of the protein nanopore-based technology for the uni-molecular detection of analytes. Subsequently, we aim to harness our findings and bring them to the next level (TRL3), in which we will demonstrate and validate the ‘proof-of-concept’ of an integrated, portable device enabling the detection of HBeAg in a variety of real-world biological samples.
Objective 1: Single-molecule study of the interactions between HBeAg and its antibody, anti-HBeAg, with the α-HL nanopore.
Objective 2: Proof-of-concept of real-time fingerprinting of HBeAg, through single-molecule stochastic sensing of the HBeAg/anti-HBeAg dimer with the α-HL nanopore.
Objective 3: Development of portable and stable, α-HL nanopore-based platform, for point-of-care HBeAg detection, superior to existing technologies in terms of throughput, affinity and selectivity.
Results and dissemination
► Conferences and scientific events
► Scientific articles
- Project Coordinator: Alexandru Ioan Cuza University of Iasi
- Project leader: Prof. Dr. Tudor LUCHIAN
- Team member: CS II Dr. Alina ASANDEI
- Team member: Conf. Dr. Loredana MEREUȚĂ
- Team member: CS III Dr. Irina ȘCHIOPU
- Partner institution: The Institute of Biochemistry-IB, Bucharest
- Team leader: CS I Dr. Norica NICHITA
- Team member: CS II Dr. Cătălin LAZĂR
- Team member: PhD student
Finance by: Ministry of Ntional Education, P2 – Increasing the competitiveness of the Romanian economy through RDI (https://uefiscdi.gov.ro)
COD: PN-III-P2-2.1-PED- 2019-0016
Project Coordinator: Alexandru Ioan Cuza University of Iasi
Partner institution:The Institute of Biochemistry-IB, Bucharest
Implementation period: 01.11.2020 – 30.04.2022
Project Manager: Prof. dr. LUCHIAN Tudor, Alexandru Ioan Cuza University of Iasi