Vitti Antonella, Nuzzaci Maria, Condelli Valentina, Piazzolla Pasquale
School of Agricultural, Forestry, Food and Environmental Sciences, University of Basilicata, Potenza, Italy.
Methods Mol Biol. 2014;1108:41-56. doi: 10.1007/978-1-62703-751-8_3.
Edible vaccines must survive digestive process and preserve the specific structure of the antigenic peptide to elicit effective immune response. The stability of a protein to digestive process can be predicted by subjecting it to the in vitro assay with simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Here, we describe the protocol of producing and using chimeric Cucumber mosaic virus (CMV) displaying Hepatitis C virus (HCV) derived peptide (R9) in double copy as an oral vaccine. Its stability after treatment with SGF and SIF and the preservation of the antigenic properties were verified by SDS-PAGE and immuno western blot techniques.
可食用疫苗必须在消化过程中存活下来，并保留抗原肽的特定结构以引发有效的免疫反应。蛋白质对消化过程的稳定性可以通过将其置于模拟胃液（SGF）和模拟肠液（SIF）的体外试验中来预测。在此，我们描述了生产和使用嵌合黄瓜花叶病毒（CMV）的方案，该病毒展示了双拷贝的丙型肝炎病毒（HCV）衍生肽（R9）作为口服疫苗。通过SDS-PAGE和免疫印迹技术验证了其在SGF和SIF处理后的稳定性以及抗原特性的保留情况。
