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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2013, Vol. 39 Issue (3): 351-354    DOI: 10.3785/j.issn.1008-9209.2012.02.033
动物科学     
疫苗的泡沫冷冻干燥研究
张绍志1， 施铭耀1， 陈光明1， 吕芳2， 卢宇2*
（1.浙江大学制冷与低温研究所，杭州 310027；2.江苏省农业科学院国家兽用生物制品工程技术研究中心，南京 210014）
Study on  foam freeze drying in vaccine
ZHANG Shaozhi1，SHI Mingyao1，CHEN Guangming1，LYU   Fang2，LU Yu2*
(1. Institute of Refrigeration and Cryogenic Engineering, Zhejiang University, Hangzhou 310027, China; 2. National Research Center for Veterinary Vaccine Engineering and Technology of China, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
)
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摘要： 以鸡新城疫活疫苗为例，介绍了泡沫冷冻干燥的工艺过程，并对用泡沫冷冻干燥法制得的样品进行热分析及鸡胚半数感染量(50% egg infectious dose, EID50)测试，然后与用传统冷冻干燥法生产的低毒疫苗和耐热疫苗进行对比。结果表明：经泡沫冷冻干燥工艺处理的疫苗与低毒疫苗、耐热疫苗的玻璃化转变温度分别为33.9 ℃、30.2 ℃和40.1 ℃；泡沫冷冻干燥疫苗和耐热疫苗的耐老化试验下降滴度分别为1.1和0.8。表明用泡沫冷冻干燥法制得的疫苗在耐热性能上有其优越性，值得进一步深入研究。  
Abstract: Freeze-drying has been widely used in the production of pharmaceuticals, vaccines and high-valued foods. Foam freeze-drying is a new type of drying technology, in which the sample is foamed and frozen before drying. Compared with traditional freeze-drying method, the new method can reduce operation time and cost significantly. Some foreign researchers have used this method to produce heat-resistant vaccines which can endure higher temperature and remain effective after a longer storage in refrigerator. Up to now, little research about foam freeze-drying has been reported domestically. In this study,
newcastle disease vaccine was used as an example to illustrate the procedure of foam freeze-drying method and to test its capability of producing heat-resistant vaccine. The ingredient of the foam freeze drying (FFD) sample included  w(trehalose)=10%,  w(hydrolyzed gelatin)=5%, w(pluronic F-108)=3%,
w(arginine)=2%, 25 mmol/L  phosphate buffer (pH=7.2). The sample was prepared through the following procedure: 1) One milliliter   sample was placed on the plate of the freeze dryer and cooled to 15 ℃; 2) then the pressure was lowered to 5 Pa and foaming began; 3) after one hour the plate temperature was increased up to 31 ℃ at a velocity of 0.7 ℃/min and kept for another 48 hours. The samples made by foam freeze drying were analyzed with thermal analysis and aging test, and were compared with live attenuated samples and heat-resistant samples prepared by traditional freeze drying. The latter two kinds of sample were bought from market (Tianbang Biotechnology Corporation, Nanjing, China). Thermal analysis was done with a differential scanning calorimetry (DSC 200 F3 Maia, NETZSCH, Germany). The procedure was as follows: 1) the sample was cooled to -20 ℃ at a rate of 10 ℃/min and held for 3 minutes; 2) the sample was heated to 80 ℃ at the rate of 10 ℃/min and held for 3 minutes; 3) the sample was cooled to room temperature at the rate of 10 ℃/min. Aging test was carried out by placing the sample at 37 ℃ for 10 days and thereafter measuring its EID50. EID50 was determined by vaccination with chick embryo and Reed-Muench calculation method. The appearance of the FFD  sample was like sponge, totally different from the flat biscuit-like sample made by traditional freeze drying method. The average glass transition temperature of FFD vaccine was 33.9  ℃, higher than 30.2 ℃ of live attenuated vaccine, but lower than 40.1 ℃ of heat-resistant vaccine. This means that  the heat resistance ability of FFD vaccine is physically better than that of live attenuated vaccine. After aging test the titers decreased for FFD vaccine was 1.1, which was a little higher than 0.8 for heat-resistant vaccine. The latter result was in consistent with the former result. It is clear that FFD vaccine has  its own superiority on heat resistance ability, although the formula and operation parameters still need to be improved. Considering its obvious economic prospect, it is worth to carry out further work on the study of FFD vaccine.
出版日期: 2013-05-20
CLC:  R 392  
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卢宇2*
张绍志1
施铭耀1
陈光明1
吕芳2

引用本文:

张绍志1， 施铭耀1， 陈光明1， 吕芳2， 卢宇2*. 疫苗的泡沫冷冻干燥研究[J]. 浙江大学学报(农业与生命科学版), 2013, 39(3): 351-354.

ZHANG Shaozhi1，SHI Mingyao1，CHEN Guangming1，LYU Fang2，LU Yu2*. Study on  foam freeze drying in vaccine. Journal of Zhejiang University (Agriculture and Life Sciences), 2013, 39(3): 351-354.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.02.033    或    http://www.zjujournals.com/agr/CN/Y2013/V39/I3/351

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