Affiliation:
1. Stavropol Antiplague Institute
Abstract
Liquid erythrocyte diagnostic preparations have a practical disadvantage; i.e., long-distance transportation involving possible non-compliance with cold-chain requirements may result in a complete loss of biological activity. A lyophilisation technology is necessary to ensure that the preparations retain their original properties for a long time. The aim of the work was to develop a protective medium and conditions for lyophilisation to stabilise the erythrocyte diagnostic preparation of tularaemia immunoglobulin. Materials and methods: Gelatin, thiourea, trehalose, sucrose, dextran, and Tween 80 were used as excipients for protective media. The authors used nine strains of homologous and heterologous microorganisms of different genera and species to control the lyophilised diagnostic preparation sensitivity and specificity. Evaluation of the main stability-related quality attributes (appearance of the dried preparation, loss on drying, solubility, appearance after reconstitution, appearance after settling, sensitivity, specificity) considered the temperatures specific to the climatic zones where the in vitro diagnostics is intended to be marketed and used. Results: The authors developed protective stabilising media with different compositions, used them in freeze-drying of the preparation and carried out control testing. The most promising was the lyophilisation medium containing a smaller amount of ingredients —6% of dextran, 0.06% of Tween 80 and up to 0.01% of sodium azide—as it was the simplest one to prepare and ensured complete preservation of the quality attributes. The authors carried out practical evaluation of lyophilisation procedures, and the 12–14-hour procedure proved to be the most cost-effective. Conclusions: The results of long-term, or real time, and accelerated stability testing of the lyophilised diagnostic preparation demonstrated the possibility of two-year storage at a labelled temperature of 2–8 °C, as well as at elevated and low temperatures of 30±2 °С and –18 °С, respectively. The tests showed no negative effects of the temperatures on the controlled quality attributes.
Subject
General Earth and Planetary Sciences,General Environmental Science
Reference23 articles.
1. Iraklionova NS, Sysuev EB, Mas ES. Natural foci of dangerous and especially dangerous pathogens of infectious diseases. Tularemia. Uspekhi sovremennogo estestvoznaniya = Advances in Current Natural Sciences. 2013;9:118–9 (In Russ.)
2. Kudryavtseva TYu, Popov VP, Mokrievich AN, Kulikalova ES, Kholin AV, Mazepa AV, et al. Epizootiological and epidemiological situation on tularemia in Russia in 2020, the forecast for 2021. Problemy osobo opasnykh infektsii = Problems of Particularly Dangerous Infections. 2021;(1):32–42 (In Russ.) https://doi.org/10.21055/0370-1069-2021-1-32-42
3. Startseva OL, Kurcheva SA. Shelf life prediction of diagnostic fluorescent tularemia dry immunoglobulins. Zdorov`e naseleniya i sreda obitaniya – ZNiSO = Public Health and Life Environment – PH&LE. 2019;(2):56–60 (In Russ.)
4. Tyumenceva IS, Afanasev EN, Alieva EV, Kurcheva SA, Garkusha YY. Antigens and antisera of F. tularensis: on the issue of tularemia immunodiagnosis. Meditsinskiy vestnik Severnogo Kavkaza = Medical News of North Caucasus. 2012;1:49–52 (In Russ.)
5. Zharnikova IV, Zhdanova EV, Zharnikova TV, Startseva OL, Kurcheva SA, Geogjayan AS, et al. Comparative characteristics of biotechnology for the production of erythrocyte and latex diagnosticums to identify the causative agent of tularemia. Vestnik biotekhnologii i fiziko-khimicheskoy biologii im. Yu.A. Ovchinnikova = Yu.A. Ovchinnikov Bulletin of Biotechnology and Physical and Chemical Biology. 2019;15(4):27–31 (In Russ.)