用蒸馏水提取粘膜抗体,随后应用于RSV中和试验。

K F Hu, B Morein, M Merza
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引用次数: 2

摘要

病毒中和(VN)是评估RSV疫苗的重要功能测试,也包括考虑感染途径的呼吸道粘膜分泌。在我们之前的研究中,RSV ELISA采用了Bergquist等人描述的免疫球蛋白提取方法,但由于提取抗体所用的2%皂苷溶液具有细胞毒性,因此不适合进行病毒中和试验。为了克服这一问题,本研究测试了几种溶剂包括蒸馏水提取免疫球蛋白的能力。采用ELISA法对提取物中的抗体进行评价和比较。蒸馏水提取总IgA、RSV特异性IgA和IgG的效率与2%皂苷溶液相同。更重要的是,随后获得的器官提取物可用于病毒中和试验,而不会对细胞培养产生不利影响。因此,在同时需要进行ELISA和病毒中和试验的情况下,最终选择蒸馏水作为提取粘膜器官免疫球蛋白的溶剂。
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Using distilled water for the extraction of mucosal antibodies and the subsequent application in RSV neutralization test.

Virus neutralization (VN) is an important functional test for evaluating RSV vaccines, also encompassing in mucosal secretion of the respiratory tract considering the infection route. In our previous study, an immunoglobin extraction method described by Bergquist et al. was adopted for RSV ELISA, but it was not suitable for virus neutralization test due to the cell toxicity of the 2% saponin solution used for the antibody extraction. In order to overcome this problem, several solvents including distilled water were tested in the present study for the capacity to extract immunogloblins. Antibodies in the extracts were evaluated and compared by ELISA. Distilled water was as efficient as the 2% saponin solution for extraction of total IgA, RSV specific IgA and IgG. More importantly, the organ extracts obtained subsequently could be used for virus neutralization test without causing adverse effect on the cell culture. Therefore, distilled water was finally chosen as the solvent for immunoglobulin extraction from mucosal organs when both ELISA and virus neutralization test are required.

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