Our laboratory provides modern analytical services to biotechnology, chemical, diagnostic, medical, food and pharmaceutical companies. Our investigations meet the highest international standards, the applied state-of-the-art facilities are continuously updated. Wide range of sample quantities should be investigated. Analytical analyses of very low sample volume (hundered nanoliters, few microliters) even a single cell are feasible by different nanoLC coupled tandem mass spectrometry (nanolC-MS/MS) or matrix-assisted laser desorption tandem mass spectrometry (MALDI TOF/TOF MS).
Method development for bioanalytical research
We have experience in method development and optimization of high throughput precise sample pretreatment techniques and development of validated, qualitative and quantitative separation techniques (gas chromatography, liquid chromatography, electropheresis etc.) for bioactive compounds. Our laboratory can provide complex lipidomic, metabolic and proteomic profiling of different biological and clinical samples such as blood, serum, plasma, tears, saliva, urine, tissue biopsies, tissue sections etc. We have experience in the arrangement of animal and human clinical studies from the ethical issues to data interpretation and biostatistical analysis.
Biochemical arrays based on immunoreactions
Immunoassay is a well known and widely used bioanalytical method based on the reaction of antigen and antibody. These techniques are being used in the field of biochemistry, pharmaceutical research, therapeutic treatments, and environmental science to determine the quality, quantity or functional activity of a target compound. The method is characterized by high sensitivity, high throughput and specificity.
The immunoassay can be designed in different ways depending on the biological problem such as competitive, sandwich, quantitative, qualitative assays should be distinguished. The method allows a quantitative or qualitative analyses of one or more analytes simultaneously from a single sample.
In our laboratory most commonly used immunoassay based techniques are ELISA and RIA. These methods are very sensitive, specific technologies for both qualitative and quantitative analyses. The presence as well as the expression levels of bioactive proteins and peptides were determined from different biological matrices (serum, plasma, urine, saliva, tears, tissue sections etc.).
Proteomic study of head neck squamous cell carcinoma
Head and neck squamous cell carcinoma (HNSCC) includes a diverse group of tumors from the upper aerodigestive tract with high morbidity and mortality. Additionally, the heterogeneous microenvironment of the cancer cells, characterized by chronic inflammation and consisting of various components including neostroma, growing blood vessels and infiltrating immune cells, is of a vital importance in cancer development and behavior. Because of this cellular and molecular diversity no reliable clinical markers were validated to diagnose the early stage HNSCC.
In this study, we used MALDI imaging mass spectrometry for molecular profiling of S100A8 and S100A9 proteins from tissue samples and human saliva. The imaging mass spectrometry results were validated by immunohistochemistry and SDS-PAGE based MALDI TOF/TOF MS proteomic investigations.
Our results showed that S100A8 and A9 proteins are presented in human saliva as potential biomarkers in case of HNSCC. Moreover, the overexpression of S100A8/A9 proteins is localized in the cancer cells and in the supportive stromal regions and is not presented in the healthy tissue areas.
The upregulation of calgranulins in the neoplastic and hyperplastic epithelium as well as their salivary secretion suggest that the S100A8 and S100A9 proteins are potential biomarkers for very early stage tumorigenesis in HNSCC.
Early stage embryonic development
Human infertility is a global problem with significant social and economic impact. Successful pregnancy is a complex process that comprises unique events, including fertilization, implantation, decidualization, placentation and finally birth. Nowadays, the assisted reproduction technologies (ART) have provided remarkable impact for successful pregnancy after in-vitro fertilization. However, these methods are associated with relatively low clinical pregnancy rate of approx. 30% per transfer.
The receptive phase of uterus is marked by structural and functional maturation of endometrium. This is a limited time span when the blastocyst competency is superimposed on the receptive endometrium. It is a well known fact, that the lipid metabolism and signaling of early stage pregnancy are of a vital importance in successful embryogenesis. However, the embryo-maternal molecular communication is not well understood, nor it is understood whether the low take home baby outcome of IVF is the result of some abnormal lipid networking.
Matrix-assisted laser desorption ionization (MALDI) imaging mass spectrometry (IMS) is a new developed technique that enables the evaluation of molecular signals direct in situ from the tissue surface or thin sections. MALDI IMS is a label-free technique with the ability to visualize the distribution of even hundreds of biomolecules in a single measurement, maintaining the morphological integrity of the intact tissue by avoiding homogenization. Although previous IMS studies have provided outstanding spatiotemporal profiles of phospholipid and protein changes during the embryo implantation, the discovery of the in situ local molecular alterations during the early stage embryogenesis needs further investigations.
In this study we discovered the lipid distributions during the early stage development of natural and transferred embryos.