In the first step, 48 of 52 components of our in-house medium were divided into three groups according to their characteristics.
This study evaluates 3 selective media (Pylori agar [bioMérieux, France], BD Helicobacter agar, modified [Becton Dickinson, USA], and an in-house medium) designed for Helicobacter pylori isolation.
This interaction was most pronounced for HzSNPV, which experienced a dramatic decline in biological activity when switched from its original Excell 401 medium to the in-house VPM medium.
An overnight grown YPG culture was used to inoculate shake flask cultivations in 500 mL baffled flasks with a working volume of 100 mL: different in-house developed medium variants based on SYN6 medium were tested.
Further optimization of the process included the use of an in-house developed medium containing both undefined supplements (peptone and yeast extract) and defined components (components of SYN6 medium).
These included the use of the in-house developed medium SYN6.46d, the 'constant feeding mode' and the optimal FS composition.
This was further corroborated by testing a chemically defined in-house prototype medium in which up to three-fold higher amino acid concentrations were used.
For all bioreactor studies, cells were grown in an in-house developed medium supplemented with 2% (w/v) glycerol as a carbon source.
For 24-DWP production assay, cells were seeded as high as 1 × 10 c/mL with a daily supplement of fresh Amgen in-house production medium.
For the initial dosage and time course study, cells were cultured in Amgen in-house subculture medium in 24-deep well plate (DWP) (Corning, NY) for 5 days with the seeding density as 5 × 10 c/mL.
Using the in-house developed medium SYN6.46d for fermentation, we performed fed-batch cultivations in 300 mL DASGIP bioreactors in order to ascertain the optimum composition of the 'feeding solution' (FS).
