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用于口服乙肝疫苗的含乙肝表面抗原植物组织的冻干处理。

Freeze-drying of plant tissue containing HBV surface antigen for the oral vaccine against hepatitis B.

作者信息

Czyż Marcin, Dembczyński Radosław, Marecik Roman, Wojas-Turek Justyna, Milczarek Magdalena, Pajtasz-Piasecka Elżbieta, Wietrzyk Joanna, Pniewski Tomasz

机构信息

Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, 60-479 Poznań, Poland.

Poznań University of Life Sciences, Wojska Polskiego 28, 60-995 Poznań, Poland.

出版信息

Biomed Res Int. 2014;2014:485689. doi: 10.1155/2014/485689. Epub 2014 Oct 12.

DOI:10.1155/2014/485689
PMID:25371900
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4209752/
Abstract

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.

摘要

本研究的目的是开发一种冷冻干燥方案，以促进含有乙型肝炎病毒小表面抗原（S-HBsAg）的植物材料的成功处理，同时保留其病毒样颗粒（VLP）结构和免疫原性。研究了在生菜叶组织中对该抗原进行冷冻干燥，而无需任何分离或纯化步骤。对每个工艺步骤进行了连续评估，并应用了最佳参数。测试了几种干燥曲线和辅料。一次干燥在20°C下进行20小时，二次干燥在22°C下进行2小时的干燥曲线以及蔗糖在冷冻干燥过程中对S-HBsAg表现出有效的稳定作用。还分析了冷冻速率和后处理残留水分作为影响S-HBsAg保存的重要因素。该工艺具有可重复性，得到的产品中VLP含量高达200μg/g干重。对VLP和总抗原的检测以及动物免疫试验证实了冻干粉末中S-HBsAg的抗原性和免疫原性得以保留。长期稳定性测试表明，储存的冻干产品在4°C下可稳定保存一年，但在较高温度下会降解。因此，已建立了高效冷冻干燥工艺的基础，并获得了一种适用于口服植物源乙肝疫苗的合适半成品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/bf76f216f753/BMRI2014-485689.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/7e710b6da4a8/BMRI2014-485689.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/610cd362e6c3/BMRI2014-485689.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/991cbe4e1d1f/BMRI2014-485689.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/2bc722a3df68/BMRI2014-485689.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/4dfdacb6b0af/BMRI2014-485689.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/fb7d1f6aad39/BMRI2014-485689.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/bf76f216f753/BMRI2014-485689.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/7e710b6da4a8/BMRI2014-485689.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/610cd362e6c3/BMRI2014-485689.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/991cbe4e1d1f/BMRI2014-485689.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/2bc722a3df68/BMRI2014-485689.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/4dfdacb6b0af/BMRI2014-485689.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/fb7d1f6aad39/BMRI2014-485689.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8ee/4209752/bf76f216f753/BMRI2014-485689.007.jpg

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