Freeze-drying of plant tissue containing HBV surface antigen for the oral vaccine against hepatitis B.
Freeze-drying of plant tissue containing HBV surface antigen for the oral vaccine against hepatitis B.
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DOI：
10.1155/2014/485689
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发表时间：
2014
影响因子：
--
通讯作者：
Pniewski T
中科院分区：
文献类型：
--
作者：
Czyż M;Dembczyński R;Marecik R;Wojas-Turek J;Milczarek M;Pajtasz-Piasecka E;Wietrzyk J;Pniewski T
本研究的目的是开发一种冷冻干燥方案，以促进成功的处理含有小表面抗原的B型肝炎病毒（S-HBsAg）的植物材料，同时保留其VLP结构和免疫原性。研究了莴苣叶组织中抗原的冷冻干燥，没有任何分离或纯化步骤。对每个工艺步骤进行连续评价，并采用最佳参数。测试了几种干燥曲线和辅料。20 ° C干燥20 h（初次干燥）和22 ° C干燥2 h（二次干燥）以及蔗糖的特性表明，在冷冻干燥过程中，S-HBsAg具有有效的稳定性。冷冻速度和后处理残留水分也是影响S-HBsAg保存的重要因素。该工艺具有重现性，并提供了VLP含量高达200 µ g/g DW的产品。VLP和总抗原的测定以及动物免疫试验证实了冻干粉中S-HBsAg的抗原性和免疫原性的保留。长期稳定性测试显示，储存的冻干产品在4 ° C下稳定一年，但在高温下降解。结果表明，该冻干工艺的可行性和有效性，为研制乙肝口服植物源疫苗奠定了基础。
The aim of this study was to develop a freeze-drying protocol facilitating successful processing of plant material containing the small surface antigen of hepatitis B virus (S-HBsAg) while preserving its VLP structure and immunogenicity. Freeze-drying of the antigen in lettuce leaf tissue, without any isolation or purification step, was investigated. Each process step was consecutively evaluated and the best parameters were applied. Several drying profiles and excipients were tested. The profile of 20°C for 20 h for primary and 22°C for 2 h for secondary drying as well as sucrose expressed efficient stabilisation of S-HBsAg during freeze-drying. Freezing rate and postprocess residual moisture were also analysed as important factors affecting S-HBsAg preservation. The process was reproducible and provided a product with VLP content up to 200 µg/g DW. Assays for VLPs and total antigen together with animal immunisation trials confirmed preservation of antigenicity and immunogenicity of S-HBsAg in freeze-dried powder. Long-term stability tests revealed that the stored freeze-dried product was stable at 4°C for one year, but degraded at elevated temperatures. As a result, a basis for an efficient freeze-drying process has been established and a suitable semiproduct for oral plant-derived vaccine against HBV was obtained.
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影响因子：
--
作者：
Michel, M. -L.;Tiollais, P.
通讯作者：
Tiollais, P.
影响因子：
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作者：
Kew, M. C.
通讯作者：
Kew, M. C.
影响因子：
5.8
作者：
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通讯作者：
Akers, MJ
影响因子：
3.8
作者：
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通讯作者：
Chen, DX
影响因子：
5.5
作者：
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通讯作者：
Howard, John A.