The main goal of this project is to develop an innovative fusion protein (immunotoxin) for liver cancer treatment, with focus to Hepatocellular carcinoma (HCC). Liver cancer, primarily HCC, is the fourth leading cause of death from cancer. Proposed immunotoxins will be directed to the highly selective antigens on the tumor cells. The project assumes usage of designed fusion proteins comprised of highly selective cancer antibody fragments attached to cytotoxic domains. We plan to create immunotoxins containing cytotoxic moieties derived from catalytic domains of Pseudomonas aeruginosa exotoxin A and diphteria toxin. One of the innovative aspects of the project is the change in immunotoxins subcellular route within human cells. This modification will be a key factor regulating an immunotoxin activity dependently on cell proliferation rate, thus making toxin even more selective anti-cancer drug. The project is conducted in collaboration with the Warsaw University (CeNT) and the Institute of Medical Biology of PAS. As a result, innovative immunotoxins will be patented and commercialized in the further.

Genome scale analysis of the immune response against pathogenic micro-organisms; identification of diagnostic markers, vaccine candidates and development of an integrated micro array platform for clinical investigations.
The genome sequences of microbial organisms responsible for diseases of world-wide medical importance have been sequenced or will be available in the near future. Technologies for producing large numbers of proteins have been developed and high-throughput assays such as protein micro arrays have been clinically validated for detecting the presence of antibodies, in serum, directed against microbial antigens. These achievements offer the opportunity of investigating the natural immune response against the whole proteome of a variety of micro-organisms. Powerful combinations of genomic information, molecular tools and immunological assays are becoming available to help identify the antigens that function as targets of protective immunity or could be used as markers for serodiagnosis. We propose here to identify in micro-organisms of great medical relevance (M. pneumoniae, C. pneumoniae, L. pneumophila, coronavirus spp and P. falciparum), a large collection of surface and secreted proteins as well as putative endotoxins. This protein repertoire will be produced as recombinant molecules or as sets of overlapping synthetic peptides and printed on array slides. The serum reactivity of groups of individuals with proven history of exposure to the selected micro-organisms will be analysed against the arrayed proteins to identify diagnostic markers and correlates of protection.
This project will significantly expand the SMEs bank of Intellectual Property and contribute to expertise within the RTDs. It is anticipated that the proposed work in high throughput protein expression, software analysis, surface peptides synthesis, protein and peptide surface capture, and array reader instrumentation will create an integrated platform of great commercial and research value. Finally it will contribute to unravelling how the humoral immune response interacts with the microbial proteomes thus filling the gap between genomic data and development of novel vaccines and diagnostic tools.

Chirality is a key factor in the efficacy of many drugs and the production of single enantiomers of chiral intermediates has therefore become increasingly important. Biocatalysis offers high enantioselectivity and regioselectivity in chiral synthesis through enzyme-catalyzed reactions and thus has an important advantage over chemical synthesis. Molecular genomic data is an unprecedented resource of enzymes for biocatalysis, but rational and effective methodologies must be established to realize the full potential of these resources. This project will focus on the discovery of novel enzymes, from both public and proprietary eubacterial genomes, in particular novel alcohol dehydrogenases, cytochrome P450 monooxygenases and amino acid modifying enzymes for use in established and innovative processes for chiral synthesis.
The DataGenome project extends from genome analysis, through cloning, expression, enzyme production, screening and protein engineering, to the enzymatic production of chiral biomolecules. The design of the project takes advantage of broad funnel-approach starting with innovative data-mining and processing of a large number of genes to ensure high flow-through in the process and rational selection of best enzyme candidates. The specific combination of expertise and design of the research project is aimed at high success-rate for the development of successful biocatalysts. Emphasis will be put on effective bioinformatics analysis to minimize the requirement for the more laborious “wet chemistry” analysis as well as development of optimized vector-host systems for efficient gene expression and enzyme production. Rational protein engineering or directed molecular evolution will be employed in order to obtain more robust variants, new substrate preferences or enhanced enantiomeric selectivity. Selected enzymes will be tested in existing and/or novel biocatalytic processes for production of chiral pharmaceutical intermediates with applications in therapeutic areas including AIDS, cancer and Alzheimer’s disease.

A realization of this project will result in creation of the innovative anticancer “pro-pharmaceuticals” characterized by high effectivity and selectivity with accompanying minimalized side effects of the foreseen therapy. Research will be focused on designing, obtaining and in vitro validation of recombinant proteins called immunoendotoxins able to selectively binding, internalization and cancerous cells killing. The most important element of each immunoendotoxin will be an effector domain of a human origin thus minimalizing possible immunogenicity of the proteins. Low immunogenicity in association with known benefits of immunotoxins-based strategies will result in very potent anticancer therapeuticals.

A High-throughput and universal system for DNA removal from biological samples

The project aims to create and test an innovative system for DNA removal from biological samples. Obtained DNase will have properties allowing for easy removal or inactivation after the reactions and enabling its use in solutions with high salt concentrations. Thus, the result of the project is a response to a market demand for easy to use and efficient DNase. The product will have an application in many areas of life-science sector in the purification of RNA and proteins, as well as in in vitro expression and cell cultures. The end results of the project will be the complete protocols of industrial preparation and utilization of nuclease-purification system.

The project is co-founded by the European Union through the Smart Growth Operational Programme 2014-2020