Research project is (co) funded by the Slovenian Research Agency.
UL Member: Faculty of Mathematics and Physics
Project: Biopharmaceuticals: sensor for aggregation of protein particles based on liquid crystals
Period: 1.5.2017 - 30.4.2020
Range per year: 1,03 FTE, category D
Head: Miha Ravnik
Research activity: Natural sciences and mathematics
Research Organisations: link on SICRIS
Researchers: link on SICRIS
Citations for bibliographic records: link on SICRIS
Controlling and understanding protein aggregation is today an open fundamental and applied challenge in the development and production of proteins as biopharmaceuticals, affecting their functionality, safety, purification, stability of solutions, packaging, and transport methods. The aggregation of proteins typically depends not only on the exact molecular structure of the proteins but also on multiple (micro-) environmental parameters, including temperature, pH, buffers, surfactants, stabilisers and tonicifiers, which from biopharmaceutics perspective, requires multiple screenings of diverse protein systems before final solutions -the biological drugs- can be developed and sent to market. However, there exist very few robust and efficient methods for screening of aggregation over a broad range of proteins and over broad scale of aggregate sizes, where especially detection of subvisible protein aggregate particles is a major challenge. Therefore, developing such technology and boost fundamental knowledge in this direction would be of major advantage to the field of biopharmaceuticals. Here, the main goal of this soft matter applied physics proposal is to develop an entirely new line of sensors for protein aggregation based on liquid crystalline fluids and to explore the role of interfaces on protein aggregation, as of direct relevance to the biopharmaceutical industry. The core mechanism of this proposal will be the performance of liquid crystalline fluids as optical detector materials where the sensing will be based on the selective absorption of proteins and/or their aggregates at the protein solution-liquid crystal interface. The presence of the aggregates will result in the changes in the molecular order of the liquid crystalline fluids, which will then be detected with various non-contact optical methods. Such approach will create highly-sensitive and (~ms) fast-responsive optical sensors for aggregation with the sought-for window for detection ranging from visible to deeply sub-visible aggregate particles. Of particular scientific relevance and conceptual advance will be questions on: (i) the role of geometry and topology of the interfaces on the protein aggregation, (ii) the effect of interfaces on the size and characteristics of aggregates, as well as the surface aggregation dynamics, and (iii) the use of light and/or plasmonics as possible mechanism for the control and even reverse of the aggregation. Methodologically, the project will develop new approaches for the detection of protein aggregates at the interfaces, based on polarisation, fluorescence, and STED microscopy as well as optical spectroscopy, which are all state-of-the-art approaches in fundamental and applied challenges of modern biotechnology and sciences in general. The project will be realised by the soft matter group in Ljubljana and the biopharmaceuticals group at company Lek d.d., part or Novartis, the industrial partner of this proposal, which are both world strong groups in their fields distinguished by their joint theoretical and experimental work and high scientific standards; e.g. proposal-related publications include PNAS 2016, Nature Comm. 2016, Sci. Rep. 2016, and Nature Phys. 2015. The socio-economic deliverables of the project are designed to yield distinct outreach, contribute to the excellence of Slovenia and abroad, and contribute new fundamental and applied knowledge to Slovenian (bio)pharmaceutical industry. Finally, this proposal is aiming at the fundamental and applied cutting edge of the state-of-the-art of biopharmaceutics, to create novel scientific and technological pathways for controlling the organisation of bio-relevant macromolecules at the microscopic and macroscopic level.
The research project is organised along three main research objectives:
- Surface sensors for protein aggregation based on liquid crystals.
- Study of surface processes of protein aggregation.
- Optimisation of sensors to specific proteins or their conformations.
This project is organised in 6 Work Packages, which Target the three research objectives. The WPs are organised with complementary theoretical and experimental studies, which are running coherently and in parallel to maximise the effectiveness of the work. The WPs are:
- Liquid crystal surface sensors for protein aggregation.
- New techniques to detect protein aggregation at interfaces.
- Study of protein aggregation at flat, curved, closed, and topological interfaces.
- Control and reverse aggregation of proteins.
- Project monitoring and management.
- Dissemination, exploitation and outreach.