Generation of Zika virus-specific recombinant antibodies

寨卡病毒(ZIVR)是一种迅速出现的传染病，与小头畸形症有关。 在巴西持续爆发的疫情中，感染母亲的孩子。ZIVR与其他黄色病毒相似，如 登革热、黄热病和西尼罗河，它们是通过蚊子传播给人类的。从服务器控制ZIVR 从公共卫生的角度和适当地处理个别患者将要求ZIVR的诊断 感染可以快速而准确地进行。因为ZIVR感染的症状与 对于其他感染，不能仅基于临床理由作出诊断，需要进行诊断化验。 不幸的是，目前还没有常规和准确的检测方法来诊断ZIVR感染。现有 化验要么局限于专门的实验室，要么给出了假阳性结果 感染了其他黄病毒。我们建议显著推进ZIVR诊断的开发 通过产生特异性识别ZIVR但不识别其他黄色病毒的抗体进行检测，并使用这些抗体 构建ZIVR抗原酶联免疫吸附试验的原型。病毒特异性抗体是感染病毒的关键试剂 最广泛使用的诊断化验类型，检测血液中病毒蛋白的存在。是这样的 化验具有快速产生结果的优势，几乎适用于任何医疗保健环境。 然而，为了准确，这种检测必须使用针对单一病毒的抗体。在这里，我们将 利用单链抗体噬菌体展示技术产生针对NS1蛋白的抗体 由ZIVR制作。我们之所以选择NS1，是因为它已被证明是其他 登革热等病毒感染。将使用的具体程序包括对小鼠进行免疫以对抗 ZIVR NS1，从这些小鼠身上获取产生抗体的B细胞，并用它们来建立一个库 表达在噬菌体(感染细菌的病毒)末端的单链抗体。这 然后将筛选噬菌体文库以寻找与ZIVR结合但不与相关病毒结合的抗体克隆 标准噬菌体展示程序。具有合适结合特性的一次抗体噬菌体克隆 经鉴定后，它们将被转化为适合用于诊断分析的全长抗体。完全没有 在这一过程的各个阶段，将对候选抗体进行测试，以验证其对ZIVR的敏感性和特异性 NS1。该项目的成功完成将导致有效的高亲和力高度可用 特异性抗ZIVR NS1抗体可用于建立敏感和特异的ZIVR诊断试验。我们 在这些研究过程中，还将开发一种适合临床测试的ZIVR NS1酶联免疫吸附试验。这将是 显著提高我们准确诊断ZIVR感染的能力，从而显著提高我们的能力 跟踪、控制和适当治疗这种疾病。

Zika Virus (ZIVR) is a very rapidly emerging infectious disease that has been associated with microcephaly in the children of infected mothers in an ongoing outbreak in Brazil. ZIVR is similar to other flavaviruses, such as Dengue, Yellow Fever, and West Nile, which are transmitted to humans by mosquitos. Controlling ZIVR from a public health perspective and appropriately managing individual patients will require that the diagnosis of ZIVR infection can be made rapidly and accurately. Because the symptoms of ZIVR infection are similar to those of other infections, the diagnosis cannot be made on clinical grounds alone and a diagnostic assay is required. Unfortunately, no routine and accurate assay for the diagnosis of ZIVR infection is currently available. Existing assays are either limited to specialized laboratories or give false positive results in patients that have been infected with other flaviviruses. We propose to significantly advance the development of a ZIVR diagnostic assay by generating antibodies that specifically recognize ZIVR, but not other flavaviruses, and using these to construct a prototype ZIVR antigen ELISA assay. Virus-specific antibodies are the key reagents required for the most widely used types of diagnostic assays, which detect the presence of viral proteins in the blood. Such assays have the advantage of yielding rapid results and being applicable to almost any health care setting. However, to be accurate, such assays must use antibodies that are specific for a single virus. Here, we will use single-chain antibody phage display technology to generate antibodies specific to the NS1 protein produced by ZIVR. We have chosen NS1 because it has proven to be an excellent diagnostic target for other viral infections such as Dengue. The specific procedures that will be used include immunizing mice against ZIVR NS1, harvesting the antibody-producing B cells from these mice, and using them to make a library of single-chain antibodies that are expressed on the tips of a bacteriophage (viruses that infect bacteria). This phage library will then be screened for antibody clones that bind to ZIVR, but not to related viruses using standard phage display procedures. Once antibody phage clones with the appropriate binding characteristics are identified, they will be converted into full-length antibodies suitable for use in a diagnostic assay. At all stages of the process, candidate antibodies will be tested to validate their sensitivity and specificity for ZIVR NS1. The successful completion of this project will result in the availability of validated high affinity highly specific anti-ZIVR NS1 Abs that can be used to construct sensitive and specific ZIVR diagnostic assays. We will also develop a ZIVR NS1 ELISA suitable for clinical testing in the course of these studies. This will markedly improve our ability to accurately diagnose ZIVR infection, thus significantly improving our capacity to track, control, and appropriately treat this disease.