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Registration Number

0225U004306, (0124U004350) , R & D reports

Title

Construction of yeast producers of granulocyte-macrophage colony-stimulating factor (GM-CSF) activating wound healing

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Селекція дріжджових трансформантів та їх аналіз на предмет продукції та локалізації ГМ-КСФ, оптимізація умов культивування отриманих продуцентів, розробка методів очистки ГМ-КСФ та перевірка активності шляхом стимуляції проліферації клітин лінії TF1.

Head

Dmytruk Kostyantyn V., Доктор біологічних наук

Registration Date

28-11-2025

Organization

Institute of Cell Biology of the National Academy of Sciences of Ukraine

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The main goal of the project is to construct GM-CSF producers based on methylotrophic yeast with "humanized" protein glycosylation. Cultivation conditions of the obtained producers will be optimized to maximize GM-CSF production. Methods for GM-CSF purification will be developed. An additional objective is the immobilization of GM-CSF on the yeast cell surface. The activity of GM-CSF and yeast displaying this growth factor on surface will be evaluated by stimulating TF1 cell line proliferation.

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Within the framework of the project, integrative vectors carrying a yeast-optimized synthetic gene of human GM-CSF, including the α-factor secretion signal and S. cerevisiae anchoring sequences, were constructed. Strains of methylotrophic yeast capable of efficiently producing secreted forms of GM-CSF, as well as strains displaying the cytokine on the cell surface, were generated. Purification protocols for recombinant GM-CSF were developed, and purified preparations of the target proteins were obtained. Both the secreted and surface-displayed forms of GM-CSF were evaluated using the human TF-1 cell line, where their biological activity was confirmed. The results demonstrate the effectiveness of the engineered yeast platform for producing bioactive forms of GM-CSF. Based on the outcomes of this project, two articles were published in Scopus-indexed Q2 journals, and a utility model application was submitted. The project results were also presented at an international conference as a poster presentation.

Product Description

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Olena V. Dmytruk
Andrii A. Sybirnyi

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2025-11-28