Аннотация
Vaccinia virus (VACV) remains an important vaccine and viral-vector platform, but production in adherent cells becomes increasingly cumbersome when capacity is expanded mainly by adding planar culture units. Serum-free cultivation can further support process standardization by reducing reliance on animal-derived components and associated batch-to-batch variability. We evaluated serum-free VACV production in Vero cells using two independently operated process configurations: a conventional 10-layer Cell Factory (6,320 cm 2 growth area) and a BelloStage™-3000 tide-motion system using BelloCell® 500A vessels containing 5.5 g of BioNOC™ II macrocarriers (approximately 15,600 cm 2 manufacturer-reported surface area). The processes were compared in terms of cell expansion, endpoint infectious virus titer, nutrient behavior, cumulative infectious virus recovery, and space-time yield (STY). In the Cell Factory process, the estimated total Vero cell number increased from 5.70 × 10 7 to 2.94 × 10 8 cells, corresponding to a 5.2-fold expansion, while cell viability remained above 90%. In the BelloStage™-3000 process, the estimated total cell number increased from 8.70 × 10 7 to 1.16 × 10 9 cells, corresponding to a 13.3-fold expansion, with viability ranging from 84% to 93%. Mean final infectious virus titers were 5.75 ± 0.25 and 6.42 ± 0.14 log₁₀ TCID₅₀/mL for the Cell Factory and BelloStage™-3000 processes, respectively. Mean cumulative infectious virus yield, calculated separately for each biological replicate, was (9.39 ± 5.19) × 10 8 TCID₅₀ per replicate for the Cell Factory process and (1.35 ± 0.40) × 10 9 TCID₅₀ per replicate for the BelloStage™-3000 process, a 1.44-fold difference between the means. Corresponding mean STY values were (3.13 ± 1.73) × 10 8 and (4.51 ± 1.32) × 10 8 TCID₅₀/L/day. Glucose generally decreased during cell expansion and virus production, with transient increases after scheduled medium replacement, whereas changes in culture medium pH were modest. Under the conditions evaluated, the BelloStage™-3000 process showed a larger estimated fold expansion of Vero cells, a numerically higher mean endpoint infectious virus titer, and moderately higher cumulative infectious virus yield and STY than the Cell Factory process. The results support further evaluation of tide-motion cultivation as a laboratory-scale platform for intensified serum-free Ospa-Vaktsina production and evaluation at larger scale.