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

A:浓缩倍数	B:添加量/%
1.0	1
1.5	3
2.0	5

Table 2 Experimental design table for optimization of concentration multiple and addition amount of BHK-21 cells supplemented medium

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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
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.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
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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