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，预防艾滋病毒感染的有效疫苗将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一种敏感的、可重复的、高通量的HIV抗体中和试验。为了自信地测量和评估疫苗设计和交付方面的适度增量改进，需要灵敏度和可重复性。需要具备高通量能力，以适应在大型临床试验中对候选疫苗进行具有成本效益和及时的评估。此外，可用于评估SIV/猕猴模型保护性免疫的中和试验将有助于选择最有希望的候选物进行大规模人体试验评估。传统的艾滋病毒抗体中和试验（即基于人工PBMC的试验）不能充分满足当前和未来加强艾滋病毒疫苗开发工作的需求。

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.