Innovative affinity-based system for blood cell separation

Efficient and reliable isolation of specific cells from complex, multi-component biological samples such as blood is prerequisite for many analytical and preparative methods and assays. Recent breakthroughs, such as novel findings related to the autoimmune disorders or leukaemia demanded isolation of specific types of blood cells, more precisely subtypes of the peripheral blood mononuclear cells. Isolation of haematopoietic stem/progenitor cells (HSPCs) from cord blood, peripheral blood apheresis or bone marrow is also of extreme interest for both the scientific and medical community. Stem cells including HSPCs are already used in cell therapies. Despite multiple developed techniques efficient and reliable separation of different cell populations from blood is still particularly challenging due to the erythrocytes and blood plasma. Systems currently providing high purity and reliability of cell isolation from blood are expensive, labour intensive (FACS), or have a low yield and throughput (MACS). Also, they demand prior labelling of the cells with specific antibodies which enable isolation of the target cells. After the separation, the target cell population is therefore fluorescently or magnetically-labelled, which may be a problem for downstream applications. Antibody-independent techniques in principle render isolation of label-free target cells, however, use of most is strongly influenced by the cell size and do not permit separation of the cells of similar sizes, which is often needed. Others, based on specific materials are limited to single cell types. To bypass this inflexibility, materials may be functionalized using wide variety of complementary molecules- peptides, aptamers or antibodies, however, specific release strategies need to be used then in order to collect the target cells. The objective of the proposed project is to create an innovative passive cell separation system, enabling easy and efficient isolation of different cell populations from peripheral and cord blood with less manipulation, higher yield and lower costs than current systems. The proposed novel system is based on the state-of-the-art affinity-based cell separation using a filter that combines inert synthetic fabric, preventing unspecific cell binding, and specific antibodies to capture the target cells. In the end we will integrate the finalized filter into an automated system reducing the chances for a human error, enabling multistep cell isolation, regulation of operation and instant monitoring of the isolation yield. Our proposal upgrades the existing solutions for passive cell isolation based on synthetic polymers and with utilisation of specific antibodies adds to them flexibility characteristic for FACS and MACS systems. This will enable a wide range of isolation of different cell types from peripheral and cord blood. At the same time, with the immobilisation of the antibodies on the carrier (filter) it retains one of the key advantages of the passive cell separation systems- unchanged surface properties of the isolated cells. The project is divided into successive work packages and a package dedicated to the project management, IP protection and dissemination of the results. Activities, timelines, milestones and expected results are defined for all the WPs, as well as the potential risks and risk management solutions. The packages will be led by excellent researchers from relevant fields. The project team from the three participating institutes- Medical Faculty Maribor, Faculty for Chemistry and Chemical Technology Maribor and partner form the industry Strip's provides all the necessary expertise for a successful implementation and finalization of the project. All the necessary equipment for the implementation of the project is available. Thereby the project is completely feasible with the funds provided within the call. The preliminary feasibility study already resulted in a patent application PTC/SI2018/050035.

The objective is to create an innovative passive cell separation system, enabling easy and efficient isolation of different cell populations from peripheral and cord blood with less manipulation, higher yield and lower costs than current systems. The proposed novel system is based on the state-of-the-art affinity-based cell separation using a filter that combines inert synthetic fabric, preventing unspecific cell binding, and specific antibodies to capture the target cells. Our proposal upgrades the existing solutions for passive cell isolation based on synthetic polymers and with utilisation of specific antibodies adds to them flexibility characteristic for FACS and MACS systems. The system thus combines advantages of the passive cell separation (unchanged surface properties of the isolated cells) and of the specific antibody-based methods (flexibility). In the end we will integrate the finalized filter into an automated system enabling multistep cell isolation, regulation of operation (i.e. fluids flow on the filter, temperature regulation) and instant monitoring of the isolation yield (cell counter). Integration into an automated system also considerably reduces risk of human error. Different cell populations that could be isolated with the new system will subsequently be useful in precise study of the contribution of individual blood cell populations in complex diseases and also in development of cell therapies. The expected lower costs and higher yield of isolations also translate into greater affordability of the "downstream" applications of the isolated cells in research and therapeutic purposes, both of which is of great significance. Our proposal has a great innovative potential on a global scale. Two patent applications are possible (the filter and the automated system). Industry’s interest for the project has already been demonstrated by the willingness of the industrial partner to co-finance and actively participate in the development.

In the implementation phase the project will enable new recruitment of highly qualified researchers both in the public and private-industrial spheres in the Eastern cohesion region. Successful completion of the project will be followed by industrial development and commercialization, which will directly affect the increase in revenues from market activity on the part of the participating public institution and increased sales on the company's side. In addition to direct financial effects, the project will consolidate the position of the University of Maribor as a development partner for the industry and of the company Strip's as a development and production partner in the field of medical devices and enable it to position well in the target markets. The project will also enable the development and production of high added value products.