Development of sandwich ELISAs that can distinguish different types of coxsackievirus A16 viral particles.
Development of sandwich ELISAs that can distinguish different types of coxsackievirus A16 viral particles.
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开发可区分不同类型柯萨奇病毒 A16 病毒颗粒的夹心 ELISA
DOI：
10.1007/s00253-016-7296-z
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Xia Ningshao
中科院分区：
文献类型：
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作者：
Ye Xiangzhong;Yang Lisheng;Jia Jizong;Han Jinle;Li Shuxuan;Liu Yajing;Xu Longfa;Zhao Huan;Chen Yixin;Li Yimin;Cheng Tong;Xia Ningshao
柯萨奇病毒A16(CA16)是手足口病的主要病原体之一。到目前为止，还没有CA16候选疫苗进入临床试验。免疫原性研究表明，不同的CA16颗粒对候选疫苗的效果有很大影响。然而，目前还没有关于不同CA16颗粒检测方法的相关报道。在本研究中，我们筛选了几种针对不同CA16病毒颗粒的单抗，并建立了几种夹心酶联免疫分析方法(ELISA)来检测不同类型的CA16病毒颗粒。仅能结合变性衣壳或空衣壳的单抗不能中和CA16。相反，能与成熟全颗粒或所有类型颗粒结合的单抗显示出明显的中和活性。用这些夹心ELISA评价了不同CA16颗粒的热稳定性。研究发现，成熟的全颗粒比其他类型的颗粒更不耐热，并且可以被高浓度的阳离子稳定。这些方法可用于辅助控制CA16疫苗的效力，并将促进CA16疫苗的发展。
Coxsackievirus A16 (CA16) is one of the major causative agents of hand, foot, and mouth disease (HFMD). No CA16 vaccine candidates have progressed to clinical trials so far. Immunogenicity studies indicated that different CA16 particles have much influence on the efficacy of a candidate vaccine. However, there are still no relevant reports on the methods of detecting different CA16 particles. In this study, we screened several monoclonal antibodies (mAbs) specific for different CA16 particles, and several sandwich enzyme-linked immunoassays (ELISAs) were developed to measure the different types of CA16 viral particles. The mAbs that could only bind denatured or empty capsids could not neutralize CA16. In contrast, the mAbs that could bind mature full particles or all types of particles showed obvious neutralizing activity. The thermal stability of different CA16 particles was evaluated using these sandwich ELISAs. The mature full particles were found to be more thermolabile than the other types of particles and could be stabilized by high concentrations of cations. These methods can be used to assist in the potency control of CA16 vaccines and will promote the development of a CA16 vaccine.