Allergies are a prevalent health issue that affects many people regularly. The prevalence of allergies varies between 20% and 40% of the population in each country. However, these numbers are underestimated because many individuals do not seek medical advice or are incorrectly diagnosed [1]. Allergy is an overactive response of the immune system to substances that are typically harmless to most people [2]. These substances, known as allergens, are molecules from animal fur, dust, pollen, etc., which can trigger allergic reactions. People with allergies can experience various symptoms, including swollen tongue, itchy and/or watery eyes, etc. Allergies could also lead to chronic conditions like eczema and allergic asthma [3]. Whole-allergen extracts from natural sources are still widely used for allergy diagnostics and immunotherapy. However, the use of natural extracts, which are difficult to standardize, is limited by many factors. A varying number of different allergens, their biological activity, and contaminants in extracts can lead to false positive or negative results and ineffective immunotherapy. Single recombinant allergen molecules could be used to overcome these problems [4].
Yeasts are commonly used for the synthesis of recombinant proteins because they combine the advantages of both prokaryotic and eukaryotic cells. Yeast can perform post-translational modifications, such as disulfide bond formation and glycosylation. Additionally, yeast cultivation is cost-effective, simple, and allows for producing recombinant proteins in high yields. Nevertheless, a significant drawback of using yeast is the hyperglycosylation of proteins, which can impact protein antigenicity. Therefore, testing different yeast species and strains can benefit recombinant protein production, as their distinct glycosylation profiles may help reduce the effects of hyperglycosylation [5].
In this study, recombinant allergen Art v 3 from Artemisia vulgaris (common mugwort) was analyzed. A. vulgaris is prevalent in Europe, North America, and Asia. During the pollen season in late summer and fall, common mugwort is one of the main causes of pollinosis [6]. In Europe, A. vulgaris accounts for approximately 10% to 14% of all cases of pollinosis [7]. Artemisia vulgaris has six different allergens (Art v 1-6), and up to 70% of individuals allergic to common mugwort can be sensitized to Art v 3. Art v 3 belongs to the family of non‐specific lipid transfer proteins. [6]. During this study, recombinant Art v 3 protein synthesis experiments were conducted using three different yeast species: Saccharomyces cerevisiae, Kluyveromyces lactis, and Pichia pastoris. The recombinant allergen proteins were purified, and primary antigenicity experiments were carried out. The research is funded by the Research Council of Lithuania (LMTLT), agreement No S-ST-24-73.