| Influenza Virus-cell Fusion Inhibition Assay
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Author:
Date:
2013-12-20
[Abstract] During viral infection to host cells, several viruses undergo the process of endocytosis and pH-dependent fusion. By fusion of viral membrane with host cellular membrane, the viral core invades to host cytoplasm. A part of monoclonal antibodies against viral membrane protein have potential to inhibit the viral fusion step. Here we describe in vitro influenza virus-cell fusion inhibition assay. The infected cells expressing viral membrane protein, such as hemagglutinin (HA), on cellular surface are incubated with monoclonal antibodies targeting viral membrane protein. Then the cells are incubated under low pH condition. If the antibody does not inhibit the fusion step, we can see multinucleated giant cells.
[摘要] 在病毒感染宿主细胞期间，几种病毒经历内吞作用和pH依赖性融合的过程。 通过病毒膜与宿主细胞膜的融合，病毒核心侵入宿主细胞质。 抗病毒膜蛋白的单克隆抗体的一部分具有抑制病毒融合步骤的潜力。 在这里我们描述了体外流感病毒 - 细胞融合抑制试验。 将表达细胞表面上的病毒膜蛋白（例如血凝素（HA））的感染细胞与靶向病毒膜蛋白的单克隆抗体温育。 然后将细胞在低pH条件下温育。 如果抗体不抑制融合步骤，我们可以看到多核巨细胞。
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| Isolation of Neutralizing Antibody
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Author:
Date:
2013-12-20
[Abstract] Use of monoclonal antibodies (MAbs) is an established laboratory strategy for characterization of specific pathogens and their antigenicity. Especially, Human MAbs (HuMAbs) with neutralizing activity against specific virus could have potential therapeutic application, and provide significant information on human epitopes that could be important for developing the next generation of universal vaccines against the virus. In addition to the classical method for murine MAb preparation, several methods for the preparation of HuMAbs have been developed. Here, we describe the development of neutralizing HuMAbs against specific virus. HuMAbs are established by fusion of the peripheral blood mononuclear cells of vaccinated volunteers or patients with the fusion partner cell line, named SPYMEG. ...
[摘要] 使用单克隆抗体（MAbs）是用于表征特定病原体及其抗原性的已建立的实验室策略。 特别地，具有针对特定病毒的中和活性的人MAb（HuMAb）可具有潜在的治疗应用，并提供对于可能对于开发下一代针对该病毒的通用疫苗重要的人表位的重要信息。 除了用于鼠MAb制备的经典方法外，已经开发了几种制备HuMAbs的方法。 在这里，我们描述中和HuMAbs针对特定病毒的发展。 HuMAbs通过将接种的志愿者或患者的外周血单核细胞与融合伴侣细胞系（称为SPYMEG）融合而建立。 然后确认每个制备的HuMAb是否可以通过体外中和测定中和特异性病毒。
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