RESUMEN
Chinese hamster ovary (CHO) cells are commonly used as "bio-machines" to pro-duce monoclonal antibodies (mAb) because of their ability to produce very complex proteins. In this study, we evaluated the effects of pine needle water extract (PNWE), pine needle ethanol extract (PNEE), and pine needle polysaccharide extract (PNPE) on the CHO cell growth, mAb production and quality using a Fed-batch culture process. PNPE maintained high VCD and viability, and the titer increase was correlated with its concentration. Three extracts effectively reduced the acidic charge variant and modulated mAb glycosylation. PNPE had the most profound effect, with G0F decreasing by 8.7% and G1Fa increasing by 6.7%. The change in the glycoform was also closely related to the PNPE concentration. This study demonstrated that PNPE could facilitate CHO cell growth, increase the mAb production, decrease acidic charge variants, and regulate mAb glycoforms. To identify the components responsible for the above changes, the sugar and flavonoid contents in the extracts were determined, and the chemical compounds were identified by LC-MS, resulting in 38 compounds identified from PNPE. Rich in sugars and flavonoids in these three extracts may be related to increased CHO cell growth and productivity, and changes in glycoforms.
Asunto(s)
Anticuerpos Monoclonales , Técnicas de Cultivo Celular por Lotes , Cricetinae , Animales , Cricetulus , Células CHO , Técnicas de Cultivo Celular por Lotes/métodosRESUMEN
Despite having a high early mechanical strength and using sulfoaluminate cement as the primary cementitious material, self-compacting repair mortar (SCRM) suffers from rapid hydration rates leading to construction time constraints. This study examined how several forms of calcium sulfate, including hemihydrate gypsum, anhydrite, and dihydrate gypsum, affected SCRM's workability, hydration process, and microstructure. The outcomes demonstrated that adding hemihydrate gypsum sped up SCRM's early hydration rate and boosted its expansion rate. For a cement with 8% hemihydrate gypsum, 6 h after adding the water, the flexural strength and compressive strength increased by 39.02% and 34.08%, respectively. The hydration rate of SCRM can be efficiently delayed by dihydrate gypsum, although the result is subpar. The material exhibited the lowest fluidity loss in 20 min, the setting time was extended, and the 28-day flexural and compressive strengths were raised by 26.56% and 28.08%, respectively, after adding 8 percent anhydrite.
RESUMEN
Acyloins are useful organic compounds with reactive adjacent hydroxyl group and carbonyl group. Current research is usually constrained to acetoin (i.e. 3-hydroxy-2-butanone) and the biological production of other acyloins was scarcely reported. In this study, two hydroxy-pentanone metabolites (3-hydroxy-2-pentanone and 2-hydroxy-3-pentanone) of Bacillus sp. H15-1 were identified by gas chromatography-mass spectrometry and authentic standards. Then the complete genome of this strain was sequenced and de novo assembled to a single circular chromosome of 4,162,101bp with a guanine-cytosine content of 46.3%, but no special genes were found for the biosynthesis of the hydroxy-pentanones. Since hydroxy-pentanones are the homologues of acetoin, the two genes alsD and alsS (encoding α-acetolactate decarboxylase and α-acetolactate synthase, respectively) responsible for acetoin formation in this strain were respectively expressed in Escherichia coli. The purified enzymes were found to be capable of transforming pyruvate and 2-oxobutanoate to the two hydroxy-pentanones. This study extends the knowledge on the biosynthesis of acyloins and provides helpful information for further utilizing Bacillus sp. H15-1 as a source of valuable acyloins.