PDF(Pubmed)
Abstract：
BACKGROUND: We developed a MARC-145 cell culture and porcine reproductive and respiratory syndrome (PRRS) vaccine production using a novel CelCradle bioreactor. CelCradle is a packed-bed bioreactor capable of both batch and perfusion culture, and the operating parameters are easy to optimize.
RESULTS: In this study, CelCradle reached a maximum cell density of 8.94 × 105 cells/mL at 5 days post-seeding when seeded at 8.60 × 104 cells/mL (doubling time = 35.52 h). Inoculation of PRRS vaccine candidate, K418DM1.1, was performed at a multiplicity of infection (MOI) of 0.01 at 5 days post-seeding, which resulted in a high viral titer of 2.04 × 108 TCID50/mL and total viral load of 1.02 × 1011 TCID50/500 mL at 2 days post-infection (dpi). The multilayer cultivation system, BioFactory culture, yielded a higher doubling time (37.14 h) and lower viral titer (i.e., 8.15 × 107 TCID50/mL) compared to the CelCradle culture. Thus, the culture medium productivity of the CelCradle culture was 2-fold higher than that of the BioFactory culture. In the animal experiment, the CelCradle-produced vaccine induced high levels of neutralizing antibodies and effectively protected pigs against homologous challenge, as shown by the significantly lower levels of viremia at 1- and 7-days post-challenge (dpc) compared to the non-vaccinated pigs.
CONCLUSIONS: Overall, this study demonstrates that the CelCradle system is an economical platform for PRRS vaccine production.
RESULTS: In this study, CelCradle reached a maximum cell density of 8.94 × 105 cells/mL at 5 days post-seeding when seeded at 8.60 × 104 cells/mL (doubling time = 35.52 h). Inoculation of PRRS vaccine candidate, K418DM1.1, was performed at a multiplicity of infection (MOI) of 0.01 at 5 days post-seeding, which resulted in a high viral titer of 2.04 × 108 TCID50/mL and total viral load of 1.02 × 1011 TCID50/500 mL at 2 days post-infection (dpi). The multilayer cultivation system, BioFactory culture, yielded a higher doubling time (37.14 h) and lower viral titer (i.e., 8.15 × 107 TCID50/mL) compared to the CelCradle culture. Thus, the culture medium productivity of the CelCradle culture was 2-fold higher than that of the BioFactory culture. In the animal experiment, the CelCradle-produced vaccine induced high levels of neutralizing antibodies and effectively protected pigs against homologous challenge, as shown by the significantly lower levels of viremia at 1- and 7-days post-challenge (dpc) compared to the non-vaccinated pigs.
CONCLUSIONS: Overall, this study demonstrates that the CelCradle system is an economical platform for PRRS vaccine production.
摘要：
背景：我们使用新型CelCradle生物反应器开发了MARC-145细胞培养和猪繁殖与呼吸综合征（PRRS）疫苗生产。CelCradle是一种填充床生物反应器，能够进行分批和灌注培养，操作参数易于优化。
结果：在这项研究中，以8.60×104个细胞/mL（倍增时间=35.52h）接种后5天，CelCradle的最大细胞密度达到8.94×105个细胞/mL。PRRS候选疫苗接种，K418DM1.1，在接种后5天以0.01的感染复数（MOI）进行，在感染后2天（dpi），导致2.04×108TCID50/mL的高病毒滴度和1.02×1011TCID50/500mL的总病毒载量。多层栽培系统，生物工厂文化，产生较高的倍增时间(37.14小时)和较低的病毒滴度(即，8.15×107TCID50/mL）与CelCradle培养物相比。因此，CelCradle培养物的培养基生产率比BioFactory培养物高2倍。在动物实验中,CelCradle生产的疫苗诱导高水平的中和抗体，并有效地保护猪免受同源攻击，与未接种疫苗的猪相比，在攻击后1天和7天（dpc）的病毒血症水平显着降低。
结论：总体而言，这项研究表明，CelCradle系统是PRRS疫苗生产的经济平台。
结果：在这项研究中，以8.60×104个细胞/mL（倍增时间=35.52h）接种后5天，CelCradle的最大细胞密度达到8.94×105个细胞/mL。PRRS候选疫苗接种，K418DM1.1，在接种后5天以0.01的感染复数（MOI）进行，在感染后2天（dpi），导致2.04×108TCID50/mL的高病毒滴度和1.02×1011TCID50/500mL的总病毒载量。多层栽培系统，生物工厂文化，产生较高的倍增时间(37.14小时)和较低的病毒滴度(即，8.15×107TCID50/mL）与CelCradle培养物相比。因此，CelCradle培养物的培养基生产率比BioFactory培养物高2倍。在动物实验中,CelCradle生产的疫苗诱导高水平的中和抗体，并有效地保护猪免受同源攻击，与未接种疫苗的猪相比，在攻击后1天和7天（dpc）的病毒血症水平显着降低。
结论：总体而言，这项研究表明，CelCradle系统是PRRS疫苗生产的经济平台。
