China Biotechnology

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Intelligent Production Process for PAT Controlled PRV Vaccine Based on Online Raman and Online Live Cell Detection

Nan WANG, Rui MIN, Xiaoping YI, Dahe ZHANG

China Biotechnology, 2024, 44(6): 53-67. DOI: 10.13523/j.cb.2312003

Fig.7 Related parameters of BHK-21 cells shake flask fed-batch culture in low glucose and glutamine concentration Control:6.0 g/L glc+0 gal+7.0 mmol/L gln;^* P<0.05,^** P< 0.01,^**** P<0.000 1

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Fig.1 Figure of feeding time calculation formula
Fig.2 Related parameters of BHK-21 cells cultured at different initial glucose concentrations in the first 12 h of culture Control:6.0 g/L glc;^* P<0.05,^** P<0.01,^*** P<0.001,^**** P< 0.000 1
Fig.3 Related parameters of BHK-21 cells cultured at different supplemental concentrations of galactose in the first 12 h of culture Control 1: 1.5 g/L glc+0 gal; Control 2: 6.0 g/L glc+0 gal; ^* P<0.05,^** P<0.01,^*** P<0.001
Fig. 4 Related parameters of BHK-21 cells cultured at different initial glutamine concentrations in the first 12 h of culture Control:6.0 g/L glc+0 gal +7.0 mmol/L gln; ^* P<0.05,^** P<0.01,^*** P<0.001,^**** P<0.000 1
Table 1 Correlation coefficient table of multivariate data analysis to determine key nutrients
Fig.5 Heat map of multivariate data analysis to determine key nutrient
Table 2 Experimental design table for optimization of concentration multiple and addition amount of BHK-21 cells supplemented medium
Fig.6 Optimization of concentration multiple and addition amount of BHK-21 cells’ supplemented medium: Response surface results of 36 h cell density (A) & 36 h specific growth rate (B)
Fig.8 Relative maximum virus titer of PRV produced by BHK-21 cells shake flask fed-batch culture in low glucose and glutamine concentrations Control:6.0 g/L glc+0 gal+7.0 mmol/L gln;^**** P<0.000 1
Fig.9 Online parameter changes of BHK-21 cells reactor manual fed-batch culture in low glucose and glutamine concentration conditions A:No feeding B:24 h feeding 1.5 times the initial medium, 5% of the total volume;The optimal initial glucose and glutamine concentrations were 1.5 g/L and 2 mmol/L, respectively and the supplemented concentration of galactose was 6 mmol/L. During the fed-batch culture, glucose and glutamine concentrations were controlled at about 0.5 g/L and 1.5 mmol/L, respectively
Fig.10 A: High glucose control B: Reactor manual feeding
Fig.11 Cell metabolism parameters of PRV produced by BHK-21 cells under different culture conditions
Fig.12 On-line Cap and virus titers of PRV produced by BHK-21 cells under online Raman detection combined with PAT feedback control and highest relative virus titer of PRV under different culture conditions ^*** P<0.001,^**** P<0.000 1
Fig.13 Off-line/on-line detection parameters of PRV produced by BHK-21 cells under different culture conditions A: Glucose concentration curve B: Glutamine concentration curve

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