A SCALABLE PHOTOCHEMICAL INACTIVATION PROCESS FOR HIV

可扩展的 HIV 光化学灭活工艺

• 批准号: 6148263
• 负责人: HAYNES W SHEPPARD
• 金额: $ 16.2万
• 依托单位: PUBLIC HEALTH FOUNDATION ENTERPRISES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6148263
• 关键词: AIDS vaccines; Animalia; HIV envelope protein gp120; active immunization; mass tissue /cell culture; photochemistry; psoralens; ultraviolet radiation; vaccine development

A traditionally successful approach to viral vaccines, whole inactivated virus (WIV), has received relatively little attention in the overall effort to develop an effective HIV/AIDS vaccine. Only one inactivated virus product (the Salk/IRC Immunogen) has been tested in humans and this product is envelope depleted and has been tested only as a therapeutic rather than a prophylactic immunogen. Since major target epitopes for antibody neutralization are in the envelope glycoprotein, it is generally considered a desirable component of a prophylactic immunogen. Thus far, however, immunization of humans with monomeric recombinant envelope glycoprotein has resulted in relatively type-specific neutralization and very little neutralization of primary isolates. The few monoclonal antibodies that demonstrate broad primary- isolate neutralization generally recognize conformation dependent epitopes. Such epitopes might be retained on whole inactivated virions. Photochemical treatment (PCT) with psoralen and UV light is a promising approach to inactivating HIV without disrupting "native" oligomeric structures. We propose to investigate a potentially scaleable process using a proprietary variant of psoralen (5-59) that lacks the in vitro mutagenicity associated with conventional psoralen methods. In addition, 5-59 inactivated products have undergone extensive safety testing in humans. The objectives of this study will be l) to investigate methods to produce and concentrate pilot-scale preparations of HIV with minimal envelope loss, 2) to confirm and Optimize a photochemical treatment process, which will use the novel psoralen 5-59, and UVA to inactivate HIV to an acceptable level while retaining gpl20 and its conformational epitopes, 3) to design/construct a novel flow PCT that is capable of treating laboratory scale quantities (5 L) of HIV culture and is capable of being scaled-up to treat production scale quantities (5000 L), 4) investigate and identify potential process steps that can be used to reduce levels of residual psoralen while maintaining native gp120 structure, 5) investigate potential interactions between PCT and other inactivation processes and determine an optimal process sequence and conditions for obtaining a double- inactivated WIV with intact gpl20, and 6) investigate animal immunogenicity of these preparations.

传统上成功的病毒疫苗方法，全灭活病毒（WIV），在开发有效的HIV/AIDS疫苗的总体努力中受到的关注相对较少。只有一种灭活病毒产品（Salk/IRC免疫原）在人体中进行了测试，该产品是包膜耗尽的，仅作为治疗性而非预防性免疫原进行了测试。由于抗体中和的主要靶表位在包膜糖蛋白中，因此通常认为其是预防性免疫原的理想组分。然而，到目前为止，用单体重组包膜糖蛋白免疫人类导致了相对类型特异性中和和非常少的初级分离株的中和。少数表现出广泛的一级分离物中和作用的单克隆抗体通常识别构象依赖性表位。这样的表位可以保留在整个灭活的病毒粒子上。用补骨脂素和紫外线进行光化学处理（PCT）是一种有前途的灭活HIV而不破坏“天然”寡聚体结构的方法。我们建议研究一种潜在的可规模化工艺，该工艺使用了一种专有的pesticen变体（5-59），该变体缺乏与传统pesticen方法相关的体外致突变性。此外，5-59个灭活产品已在人体中进行了广泛的安全性测试。本研究的目的将是1）研究以最小的包膜损失生产和浓缩中试规模的HIV制剂的方法，2）确认和优化光化学处理过程，其将使用新的peptide 5-59和UVA将HIV抑制到可接受的水平，同时保留gp 120及其构象表位，3）设计/构建新型流式PCT，其能够处理实验室规模量（5 L）的HIV培养物，并且能够扩大规模以处理生产规模量（5000 L），4）研究和鉴定可用于降低残留peptide水平同时保持天然GP 120结构的潜在工艺步骤，5）研究PCT和其他灭活过程之间的潜在相互作用，并确定用于获得具有完整gp 120的双重灭活WIV的最佳过程顺序和条件，6）研究这些制剂的动物免疫原性。