Effects of Different Emulsification Techniques on Stability of Porcine Circovirus Type 2 Inactivated Vaccine
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摘要: 为探讨乳化工艺中影响猪圆环病毒2型油包水剂型灭活疫苗的稳定性因素，以提升猪圆环病毒疫苗的质量，分别对圆环病毒抗原pH值、乳化剪切机转数、乳化时间进行筛选，并以疫苗黏度、粒径、保存温度评价疫苗稳定性。结果表明，抗原pH值为7.0~7.5时疫苗稳定性较好；剪切机转数越高，乳化时间越短，乳化效果越好，样品稳定性较高；在一定的时间内样品随乳化时间的延长，样品稳定性增强，但黏度不断增大。在本试验条件下，选择抗原pH值7.0~7.5，剪切机转数19 000 r/min，乳化时间8 min乳化的样品稳定性好，黏度合适，建议不同厂家在疫苗产业化生产时需调节抗原pH值至7.0~7.5，根据不同的乳化设备进行乳化试验摸索乳化剪机的转速及乳化时间，从而保证疫苗的稳定性。Abstract: Factors affecting the emulsion stability of the inactivated type 2 porcine circovirus vaccine were investigated. The pH, speed and time applied in the emulsification process were varied to compare the stability of the emulsion in the aspects of viscosity, particle size and storage temperature. The results showed that the emulsion was most stable at pH 7.0-7.5 with high shear and short mixing time. Within a limit, prolonged shearing would increase the stability but also raised the viscosity of the resulting emulsion. It was concluded that the optimal conditions to emulsify the vaccine were pH 7.0-7.5 at 19 000 r·min-1 for 8 min.It was recommended to keep the pH of antigen at 7.0-7.5 in the production of porcine circovirus vaccine, and determine the speed and time of emulsification according to different equipment to keep the vaccine stable.
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图 2 不同乳化时间对粒径影响分布
Figure 2. Particle size distribution as affected by emulsifying time
表 2 不同转数对疫苗稳定性的影响
Table 2. Effect of shearing speed on stability of vaccine emulsion
稳定性检测 剪切机转数/(r·min-1) 10000 13000 16000 19000 低温离心观察 +++ ++ + - 常温离心观察 - - - - 2~8℃保存观察 +++ +++ ++ - 37℃保存观察 + - - - 黏度检测/cp 37.76±0.05 37.95±0.06 38.15±0.05 38.44±0.04 
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表 3 不同乳化转数样品的粒径分布
Table 3. Particle size distribution under different shearing speed
粒径分布 乳化转速/(r·min-1) 10000 13000 16000 19000 Dv(10)/μm 0.698 0.594 0.547 0.495 Dv(50)/μm 1.140 0.928 0.842 0.756 Dv(90)/μm 1.198 1.480 1.280 1.120 注：Dv(10)n/μm代表 10%的颗粒粒径在n微米以内，Dv(50)n/μm代表 50%的颗粒粒径在n微米以内，Dv(90)n/μm代表 90%的颗粒粒径在n微米以内。表 5同。
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表 4 不同乳化时间对疫苗稳定性的影响
Table 4. Effect of emulsifying time on stability of vaccine emulsion
稳定性检测 乳化时间/min 3 5 8 10 低温离心观察 +++ + + - 常温离心观察 - - - - 2~8℃保存观察 +++ +++ ++ - 37℃保存观察 + - - - 黏度检测/cp 38.03±0.04 38.17±0.05 38.28±0.03 38.51±0.06
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表 5 不同乳化时间样品的粒径分布
Table 5. Particle size distribution under different emulsifying time
粒径分布 乳化时间/min 3 5 8 10 Dv(10)/μm 0.587 0.541 0.518 0.488 Dv(50)/μm 0.919 0.832 0.790 0.750 Dv(90)/μm 1.470 1.260 1.190 1.120
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