[1]徐悦,常晨,侯继波,等.PCV2 病毒生物矿化条件的优化[J].江苏农业学报,2017,(01):159-165.[doi:10.3969/j.issn.1000-4440.2017.01.026 ]
　XU Yue,CHANG Chen,HOU Ji-bo,et al.Optimization of biomineralization conditions of porcine circovirus type 2(PCV2) viral particles[J].,2017,(01):159-165.[doi:10.3969/j.issn.1000-4440.2017.01.026 ]
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PCV2 病毒生物矿化条件的优化()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年01期
页码:
159-165
栏目:
畜牧兽医·水产养殖
出版日期:
2017-02-28

文章信息/Info

Title:
Optimization of biomineralization conditions of porcine circovirus type 2(PCV2) viral particles
作者:
徐悦12; 常晨12; 侯继波12; 刘娅梅12
（1. 江苏省农业科学院国家兽用生物制品工程技术研究中心，江苏南京210014；2. 江苏省动物重要疫病与人兽共患病防控协同创新中心，江苏扬州225009）
Author(s):
XU Yue12; CHANG Chen12; HOU Ji-bo12; LIU Ya-mei12
(1. National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China；2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China)
关键词:
猪圆环病毒2型; 生物矿化; 磷酸钙; 浓缩纯化; 抗体水平
Keywords:
porcine circovirus type 2; biomineralization; calcium phosphate; concentration and purification; antibody titer
分类号:
S852.4+3
DOI:
10.3969/j.issn.1000-4440.2017.01.026
文献标志码:
A
摘要:
以矿化效率为评价标准，对猪圆环病毒2型（PCV2）生物矿化的反应温度、反应时间及Ca/P比、抗原初始pH、添加剂用量等条件进行优化，并通过小鼠实验检测矿化病毒的免疫原性。结果显示，生物矿化最优处理条件为初始pH 6.5，Ca/P 1.67，4 ℃矿化12 h，Mg2+/Ca2+为1.0。在该反应条件下，病毒粒子能够形成球形结构的钙磷PCV2粒子，粒径为200～500 nm。不同批次PCV2抗原矿化率皆在90%以上，且该方法可去除大量杂蛋白。小鼠免疫矿化PCV2病毒颗粒后14~28 d，抗体水平与商品苗对照组、未矿化PCV2病毒组基本相同，28 d后对照组抗体水平下降明显，而PCV2矿化颗粒组的抗体基本维持在原水平，表明PCV2矿化颗粒具有缓释抗原的作用。
Abstract:
Biomineralization is a rapid and effective method for the concentration and purification of porcine circovirus type 2 (PCV2). In this study, the conditions of PCV2 biomineralization were optimized, including reaction temperature and time, Ca/P ratio, initial pH of viral culture, and the amount of additives, to reach a high biomineralization rate. The viral culture was adjusted for an initial pH of 6.5, followed by the addition of calcium and phosphate ions (a Ca/P ratio of 1.67). Appropriate amount of Mg2+ was added for a final Mg2+/Ca2+ of 1.0. The mixture was allowed to react at 4 ℃ for 12 h. Under this optimized condition, viral particles were wrapped by calcium phosphate, forming spherical nanoparticles (CaP-PCV2) in the size between 200 and 500 nm, as indicated by electron microscopic observation. Different batches of PCV2 viruses biomineralized under this condition and yielded biomineralization rates of more than 90%. Further animal experiments revealed that the PCV2-specific antibody titer in mice injected with biomineralized PCV2 particles was similar to those in untreated PCV2 and commercialized PCV2 vaccine group 14-28 d after immunization. More importantly, the PCV2-specific antibody titer in biomineralized PCV2 group remained high until day 56, whereas the titers in both untreated PCV2 and commercialized PCV2 vaccine group started to decrease after day 28, indicating a longer duration of high antibodies of biomineralized PCV2.

参考文献/References:

[1]GRAU-ROMA L, CRISCI E, SIBILA M, et al. A proposal on porcine circovirus type 2 (PCV2) genotype definition and their relation with postweaning multisystemic wasting syndrome (PMWS) occurrence[J]. Veterinary Microbiology, 2008, 128(1/2):23-35.
[2]郭广富，曹军平，朱爱萍，等.猪圆环病毒2型泰州株的分离鉴定[J].江苏农业科学，2016，44（4）：294-295.
[3]曹东阳，王小敏，钱爱东，等．江苏省及周边地区猪圆环病毒Ⅱ型(PCV2)分子流行病学调查[J]．江苏农业学报，2016，32(2)：390-398．
[4]孙华伟，张敬峰，何孔旺，等. 2014年江苏省部分中小规模猪场PCV-2的流行病学调查[J]．山东农业科学，2015，47(4)：117-119.
[5]郎洪武，陈晓春，吴华伟，等. 猪圆环病毒病及其生物制品学的研究进展[J]. 中国兽医科学，2012，42（5）: 545-550.
[6]张许科，孙进忠，白朝勇. 猪圆环病毒2型与猪细小病毒二联灭活疫苗及其制备方法：CN 102961742A[P]2013-03-13.
[7]吕茂杰，梁武，杨保收，等. 一种二联灭活疫苗制备方法：CN 104474542A[P]2015-04-01.
[8]李倬,吴全忠,米娜,等. 一种猪圆环病毒2型纯化方法：CN 201410145472.6[P]2014-07-23.
[9]WEINER S, DOVE P M. An overview of biomineralization processes and the problem of the vital effect[J].Rev Mineral Geochem, 2003, 54(1):1-29.
[10]杨宇玲, 王广川, 唐睿康. 基于生物矿化的生物壳工程[J]. 中国科学: 化学,2014, 44(4): 601- 610.
[11]李坚, 邱坚. 生物矿化原理与木材纳米结构复合材料[J]. 林业科学, 2005, 41(1):189-193.
[12]SUBBURAMAN K, PERNODET N, KWAK S Y, et al.Templated biomineralization on self-assembled protein fibers[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(40):14672-14677.
[13]DURUCAN C, BROWN P W. Biodegradable hydroxyapatite-polymer composites[J]. Advanced Engineering Materials, 2001, 3(4):227-231.
[14]BARROUG A, KUHN L T, GERSTENFELD L C, et al. Interactions of cisplatin with calcium phosphate nanoparticles: in vitro controlled adsorption and release[J]. Journal of Orthopaedic Research, 2004, 22(4):703-708.
[15]AMRA T, KESTER M, ADAIR J H. Calcium phosphate-based composite nanoparticles in bioimaging and therapeutic delivery applications[J]. Wiley Interdisciplinary Reviews Nanomedicine & Nanobiotechnology, 2012, 4(1):96-112.
[16]HE Q, MITCHELL A R, JOHNSON S L, et al. Calcium phosphate nanoparticle adjuvant[J]. Clinical & Diagnostic Laboratory Immunology, 2000, 7(6):899-903.
[17]KUMTA P N, SFEIR C, LEE D H, et al. Nanostructured calcium phosphates for biomedical applications: novel synthesis and characterization[J]. Acta Biomaterialia, 2005, 1(1):65-83.
[18]LI Y, FENG W, YANG J, et al. In vitro synthesis and characterization of amorphous calcium phosphates with various Ca/P atomic ratios[J]. Journal of Materials Science Materials in Medicine, 2007, 18(12):2303-2308.
[19]王晓雨. 病毒壳工程的研究及其应用[D]. 杭州：浙江大学, 2011.
[20]崔福斋. 生物矿化[M]. 北京：清华大学出版社，2012：5
[21]许燕波, 钱春香, 陆兆文. 柠檬酸粗化铜离子矿化产物颗粒粒径的研究[J]. 材料导报, 2013, 27(2):92-95.
[22]COSTA D O, ALLO B A, ROBERT K, et al. Control of surface topography in biomimetic calcium phosphate coatings[J]. Langmuir the Acs Journal of Surfaces & Colloids, 2012, 28(8):3871-3880. 
[23]KOPPAD S, RAJ G D, GOPINATH V P, et al. Calcium phosphate coupled Newcastle disease vaccine elicits humoral and cell mediated immune responses in chickens[J]. Research in Veterinary Science, 2011, 91(3):384-390. 
[24]WANG G, LI X, MO L, et al. Eggshell-inspired biomineralization generates vaccines that do not require refrigeration[J]. Angew Chem Int Ed Engl, 2012, 51(42):10576-10579.

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备注/Memo

备注/Memo:
收稿日期：2016-01-04 基金项目：公益性行业（农业）科研专项项目
[201303046]；江苏省农业科技自主创新基金项目[CX（11）4037] 作者简介：徐悦（1986-），女，江苏南京人，硕士研究生，研究方向为预防兽医。（Tel）18101599119；（E-mail）xellos05@163.com 通讯作者：刘娅梅，（Tel）18551809901；（E-mail）lym613@hotmail.com
更新日期/Last Update: 2017-04-12