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We develop tools for diagnostics and treatment of socially significant diseases based on DNA-Aptamers.

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Aptamers for the diagnosis of lung cancer

Objective: to obtain DNA aptamers for the detection of lung cancer biomarkers in blood and tissue

Aptamers were selected from a DNA library of oligonucleotides using the Cell SELEX method. Postoperative tissues of patients with non-small cell lung cancer were used as a positive target for selection; adjacent tissues and whole blood of healthy people were used as a negative target.

Aptamers that specifically bind to lung cancer cells and proteins have been obtained through selection (www.sciencedirect.com). Using mass spectrometry studies, putative aptamer target proteins such as neutrophil defensin 1 and 3, vimentin, actin were identified (www.sciencedirect.com). Comparative analysis of histological sections of lung cancer tissues, adjacent tissues and lungs of healthy people showed that the obtained aptamers stain the membranes and nuclei of malignant cells and transformed vessels (www.sciencedirect.com). It was shown that the obtained aptamers can be used to detect circulating tumor cells and microemboli (www.sciencedirect.com), as well as proteins (www.nature.com, www.sciencedirect.com) in the blood and plasma of unhealthy people. 

Aptamers for intraoperative staining of brain glioblastoma

Objective: to obtain DNA aptamers capable of specifically contrasting the boundaries of brain glioblastoma in vivo

Aptamers were selected from a DNA library of oligonucleotides using the Cell SELEX method to postoperative glioblastoma tissues. Tissues of benign brain tumors and conventionally healthy brain tissues adjacent to tumor ones were used as negative targets.
The obtained aptamers specifically bound to the primary tumor of glioblastoma and did not stain healthy brain cells (Patent RU 2 654 665 C2). The putative targets of the obtained aptamers were glial fibrillary acidic protein (GFAP), tubulin alpha-1C chain (TUBA1C) and myelin basic protein (MBP) acidic protein with post-translational modifications and pancreatic lipase-related protein 3. The 3D structure of the Gli-233 aptamer was determined using small-angle scattering and computer simulation methods. Aptamers conjugated with a fluorescent Brilliant Violet 650 dye (Gli-233BV) did not show in vivo toxicity in preclinical animal studies and were used for tumor visualization during fluorescence-guided surgery in patients. The anti-GBM aptamer was conjugated with a fluorescent Brilliant Violet 650 dye (Gli-233BV) and applied on tissues during the surgical removal.

Aptamers for the detection of bacteria Salmonella typhimurium and Salmonella eneritidis

Objective: To obtain aptamers for the detection and determining the type of Salmonella typhimurium and Salmonella eneritidis

Highly specific DNA aptamers to live Salmonella typhimurium and Salmonella eneritidis were selected via the SELEX technique.

Twelve rounds of selection were performed; each comprised a positive selection step against viable S. typhimurium or S. eneritidis and a negative selection step against heat killed S. typhimurium or S. eneritidis and a mixture of related pathogens, including Salmonella enteritidis, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii to ensure the species specificity of the selected aptamers.

Based on the obtained aptamers, electrochemical sensors were made to detect salmonella (pubs.acs.org) and determine their type (pubs.acs.org). A method for determining the antibiotic resistance of salmonella S. enteritidis and S. typhimurium using DNA aptamers was developed, which is based on the ability of aptamers to bind only to living bacteria (Patent RU 2518372). It was found that a mixture of synthetic DNA aptamers to S. enteritidis and S. typhimurium inhibits the growth of bacterial colonies by more than 70% (pubs.acs.org).

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