doi:

DOI: 10.3724/SP.J.1145.2013.01064

Chinese Journal of Appplied Environmental Biology (应用与环境生物学报) 2013/19:6 PP.1064-1068

Process Optimization of Pertactin Extraction Based on Response Surface Design


Abstract:
Pertactin(PRN) is one of the most important protective antigen of acellular pertussis(aP) vaccine, and the study of its extraction has great signif icance to its yield, purity and even the quality of the aP vaccine. This research studied the critical factors which have an effect on pertactin extraction using CCF response surface model design combined with quantitative ELISA test. By two rounds of CCF RSM model optimization, the optimal parameters of pertactin extraction were determined as temperature 60 ℃, cell concentration 50 mg/mL, time 90 min, pH 8.5 and NaCl 0.15 mol/L. The robustness parameter range of PRN extraction, which was predicted by the simulation of Monte carol and verif ied by experiments designed by Plackett Burman model, were shown in this research as temperature(58-62) ℃, cell concentration(40-60) mg/mL, time(80-100) min, pH 8.45-8.55 and NaCl(0.12-0.18) mol/L. After optimization, the product of PRN in the crude extraction was increased by 50%. The results showed that the temperature has the greatest impact, followed by pH, and the other factors, such as cell concentration, time and NaCl concentration, which interact with temperature and pH, have a relatively smaller inf luence on pertactin extraction. Fig 6, Tab 3, Ref 13

Key words:pertussis,pertactin,response surface,extraction

ReleaseDate:2015-04-15 08:42:24



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