HIV-1/SIV Antibody Neutralization Assay Improvements and Database Development

描述（由申请人提供）：专家们一致认为，有效的艾滋病毒感染疫苗需要诱导有效和广泛的中和抗体反应。候选疫苗的开发和临床评价都需要灵敏、可重复和高通量的HIV抗体中和试验。需要灵敏度和再现性来自信地测量和评估疫苗设计和递送的适度增量改进。需要高通量能力以适应在大型临床试验中对候选疫苗进行成本有效和及时的评估。此外，可用于评价SIV/猕猴模型中的保护性免疫的中和试验将有助于并能够选择最有希望的候选物用于人体试验中的大规模评价。传统的HIV抗体中和试验（即基于PBMC的手动试验）不能充分满足当前和即将到来的HIV疫苗开发工作的需求。 为了支持这些重要目标，Monogram Biosciences（前身为ViroLogic，Inc.）开发了一种灵敏的中和抗体检测试剂盒（PhenoSense HIV抗体中和检测试剂盒），该试剂盒使用表达患者分离株包膜蛋白的病毒假型（见I期最终报告）。该测定允许直接测量患者血浆中的自体中和滴度，提供筛选大组单克隆抗体对相关原代菌株的中和活性的能力，并允许直接比较不同疫苗免疫原引起的抗体应答。该测定法也适用于评价猕猴模型中的SIV包膜蛋白和抗SIV抗体应答。 在本项目的第II阶段，我们计划优化Phenosense HIV抗体中和试验的性能并完成其表征。将使用定制自动化和其他规模扩大技术来增强该测定的通量和容量。将评价该试验的灵敏度、重现性和耐用性，并与传统的基于PBMC的方法进行比较。我们将开发专注于连接包膜表型和序列分析的信息数据库系统。该数据库无疑将成为疫苗设计的有用工具。Monogram Biosciences计划将PhenoSense HIV抗体中和试验和相关数据库系统商业化，以支持政府、学术和工业实验室产生的新候选疫苗的开发和临床评价。

DESCRIPTION (provided by applicant): Experts agree that an effective vaccine against HIV infection will require the induction of potent and broad neutralizing antibody responses. Both the development and clinical evaluation of vaccine candidates requires a sensitive, reproducible and high throughput HIV antibody neutralization assay. Sensitivity and reproducibility are required to confidently measure and assess modest incremental improvements in vaccine design and delivery. High throughput capability is required to accommodate the cost effective and timely evaluation of vaccine candidates in large clinical trials. In addition, a neutralization assay that can be used to evaluate protective immunity in the SIV/macaque model will aid and enable the selection of the most promising candidates for large scale evaluations in human trials. Conventional HIV Antibody Neutralization Assays (i.e. manual PBMC based assays) do not adequately address the current and upcoming demands of heightened HIV vaccine development efforts. In support of these important objectives, Monogram Biosciences (formerly ViroLogic, Inc.) has developed a sensitive neutralizing antibody assay (PhenoSense HIV Antibody Neutralization Assay) that uses viral pseudotypes that express the envelope proteins of patient-derived isolates (see Phase I final report). This assay allows for the direct measurement of autologous neutralizing titers in patient plasma, provides the ability to screen large panels of monoclonal antibodies for neutralizing activity to relevant primary strains, and allows for the direct comparison of antibody responses elicited by different vaccine immunogens. The assay has also been adapted to evaluate SIV envelope proteins and anti-SIV antibody responses in the macaque model. In phase II of this project, we intend to optimize the performance and complete the characterization of the Phenosense HIV Antibody Neutralization Assay. The throughput and capacity of this assay will be augmented using custom automation and other scale-up techniques. The sensitivity, reproducibility and robustness of the assay will be evaluated and compared to the conventional PBMC-based method. We will develop informational database systems focused on linking envelope phenotype and sequence analysis. This database will undoubtedly become a useful tool for vaccine design. Monogram Biosciences intends to commercialize the PhenoSense HIV Antibody Neutralization Assay and associated database systems to support the development and clinical evaluation of new vaccine candidates generated in government, academic, and industrial laboratories.