The experimental substantiation of the shelf-life of "Candidocyde" vaccine Because of the difficult situation with the diagnosis and therapy of candidiasis many researchers propose to use vaccines as an alternative to anti-fungal drugs for preventing and treating candidal infections. The authors have developed "Candidocyde" vaccine against candidiasis. Aim. To determine experimentally the shelf-life of the solution of "Candidocyde" vaccine. Materials and methods. "Candidocyde" vaccine was stored at two temperature regimes: in the refrigerator at the temperature of 2-8 °С and at the room temperature of (15-25 °С). Antibody titers of the solution of "Candidocyde" vaccine were assessed every three months to prevent and treat candidiasis. For this purpose the reagent kit for immunoassay detection of antibodies of class G to C. albicans using the test system ELISA "Vector-best" was applied. Results and discussion. When conducting the studies of the vaccine stored in glass vials protected from light at the temperature from 2 ºС to 8 ºС it was found that the activity of the solution was preserved for 2 years and 3 months when preventing and treating candidal infections. The titers of antibodies in animals remained at the level of 1 : 600-1 : 4000 when preventing and treating candidiasis. The positive results were recorded in all animals in the group, i.e. all animals were healthy. It indicates that the solution of "Candidocyde" vaccine is stable for 2 years. Conclusions. When studying the shelf-life of the solution of "Candidocyde" vaccine based on antigens of cells of C. albicans and C. tropicalis fungi it has been found that it is stable for 2 years of storage in glass vials protected from light at the temperature of 2-8 ºС.

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Fungi of Candida genus are the most widespread causative agent of fungal infections. Candida causes a wide range of infections: from insignificant diseases of the skin and mucous membranes to invasion processes that can practically destroys all organs. Development of a vaccine against candidiasis is the topical issue of modern pharmacy and medicine. Suspensions of cells of Candida albicans and Candida tropicalis fungi were subjected to the action of ultrasound, then filtered through a "Vladipore" membrane MFА-МА No.3 providing separation of the biological material with the size of 10 kDa and its concentration. Then prefiltration and sterilizing filtration were carried out. The resulting purified antigens of Candida albicans fungi cells with the protein concentration of 3 mg/ml and Candida tropicalis with the protein concentration of 5 mg/ml were mixed in the ratio of 1:1 using a mixer with the rotation speed of 100 rpm for 10 min. In the experiment two-month white mice with the body weight of 18-22 g were used; there were 6 animals in the control and test groups. The animals were infected intraperitoneally with the suspension of Candida albicans in the amount of 20 mln of cells and Candida tropicalis in the amount of 60 mln of cells in the volume of 1 ml. In 5 days the cell-associated antigens of Candida fungi in the volume of 0.2 ml were injected intramuscularly to mice in the upper part of the rear right paw. In 14 days the procedure was repeated. The animals of the control group were injected with the sterile 0.9% isotonic saline solution. After that in 14 days the animals were examined and the results were determined. According to the research results it has been found that the cell-associated antigens of Candida albicans and Candida tropicalis provide the therapeutic effect in 100% of animals when injected intramuscularly in the volume of 0.2 ml.

Therapeutic effect of C. albicans proteins in concentrations 1, 2, 3, 4 and 5 mg/ml has been examined in white mice. Animals were infected i.p. with suspension of C. albicans strain CCM 335–867 in the amount of 20 million cells per 1 ml volume. After 5 days and repeatedly after 14 days mice were injected s.c. in the upper part of right hind paw with proteins of Candida cells of volume 0.2 ml. 14 days after each injection, the determination of the animal body protective functions has been carried out by the titer of specific antibodies during immunoassay. According to the data obtained, it has been found that in the s.c. administration, after the first and second injection with C. albicans protein of concentration 1, 2, 3, 4 and 5 mg/ml antibody titers increased 2 times, indicating that there is no stimulation of immune protection.

In this article, different temperature regimes of the freezing-thawing technology for the disintegration of C. albicans fungal cells have been studied. The biomass of C. albicans fungal cells in 10 mL of sterile isotonic 0.9% sodium chloride solution in a Petri dish was subjected to destruction using a five-fold freeze-thaw cycle in the following temperature ranges: from (–25 ± 2) °C to (25 ± 2) °C, from (–30 ± 2) °С to (30 ± 2) °С and from (–20 ± 2) °С to (20 ± 2) °С. In order to find the optimal temperature in each case, the content of proteins, polysaccharides, and monosaccharides was determined. According to the results obtained, it has been determined that the temperature regime from (–25 ± 2) °С to (25 ± 2) °С is the most promising since it provides the maximum release of polysaccharides and proteins in the shortest possible time with minimal energy consumption.