Manufacturing of Novel Adenovirus Prime-Boost HIV-1 Vaccine

该项目的目标是为Crucell生产临床级主病毒种子库 以及用于所述临床试验的三价临床级rAd 26和rAd 5 HVR 48疫苗产品 在项目2中。Crucell Holland BV是一家中型生物技术公司，致力于 开发传染病疫苗。特别是，Crucell率先开发了rAd 基于载体的疫苗，并已开发了来自Ad血清型的载体， 人类种群。Crucell也是临床前到临床转化开发的领导者， 用于多种病原体的基于rAd载体的疫苗。特别是，Crucell开发了强大的生产能力， 和纯化方法以生产临床级rAd疫苗。Crucell还标准化了 验证释放测定，并已开发出几种包装细胞系， 生产完全符合良好生产规范（GMP）指南。候选疫苗 基于几种不同的Ad血清型的疫苗已经由Crucell制造和发布，并且最近 在美国和南非进行了埃博拉、疟疾和结核病的临床试验。 Crucell将保证rAd疫苗的统一，高质量的生产，以及严格的 遵守标准操作程序（SOP）和严格的质量控制/质量保证 (QC/QA）标准，完全符合所有相关监管机构的要求。研究级疫苗将 将提供用于兔GLP临床前毒理学研究，临床级疫苗将 为项目2中描述的I期和IIa期临床试验生产。 在我们目前的IPCAVD AI 066305项目中，我们已经开发了rAd 26和rAd 5 HVR 48 GMP 生产工艺和释放测定，已经由FDA批准。在项目3中， 我们建议利用和改进这些现有的生产工艺和分析测定， 生产和放行多价rAd 26和rAd 5 HVR 48临床疫苗产品。到 为实现这些目标，我们提出以下三个具体目标： 1.生产并放行rAd 26和rAd 5 HVR 48的临床级主病毒种子库 在GMP条件下表达最佳HIV-1 Gag-Nef、Pol和Env抗原的载体; 2.通过优化我们目前的rAd 26和rAd 5 HVR 48载体生产工艺， 收获和下游纯化步骤的载体产量;和 3.生产、纯化、释放和监测三价临床级rAd 26的稳定性， 在GMP条件下表达最佳HIV Gag-Nef、Pol和Env抗原的rAd 5 HVR 48疫苗。

The goal of this project is for Crucell to manufacture clinical-grade master virus seed stocks and trivalent, clinical-grade rAd26 and rAd5HVR48 vaccine products for the clinical trials described in Project 2. Crucell Holland BV is a medium-sized biotechnology company that is dedicated to the development of vaccines for infectious diseases. In particular, Crucell has pioneered the development of rAd vector-based vaccines and has developed vectors from Ad serotypes that have low seroprevalence in human populations. Crucell has also been a leader in the preclinical-to-clinical translational development of rAd vector-based vaccines for a variety of pathogens. In particular, Crucell has developed robust production and purification methods to manufacture clinical-grade rAd vaccines. Crucell has also standardized and validated release assays and has developed several packaging cell lines that support pharmaceutical-scale manufacturing in full compliance with Good Manufacturing Practice (GMP) guidelines. Vaccine candidates based on several different Ad serotypes have been manufactured and released by Crucell and have recently entered clinical trials for Ebola, malaria, and tuberculosis in both the United States and South Africa. Crucell will guarantee uniform, high quality manufacturing of the rAd vaccines as well as strict adherence to Standard Operating Procedures (SOPs) and rigorous quality control / quality assurance (QC/QA) standards in full compliance with all pertinent regulatory authorities. Research-grade vaccines will be provided for the GLP preclinical toxicology studies in rabbits, and clinical-grade vaccines will be manufactured for the phase 1 and phase 2a clinical trials described in Project 2. In our current IPCAVD AI066305 program, we have developed rAd26 and rAd5HVR48 GMP manufacturing processes and release assays that have already been approved by the FDA. In Project 3, we propose to utilize and to improve these existing production processes and analytical assays to manufacture and release the multivalent rAd26 and rAd5HVR48 clinical vaccine products. To accomplish these goals, we propose the following three Specific Aims: 1. To manufacture and release clinical-grade master virus seed stocks for rAd26 and rAd5HVR48 vectors expressing the optimal HIV-1 Gag-Nef, Pol, and Env antigens under GMP conditions; 2. To improve our current manufacturing processes for rAd26 and rAd5HVR48 vectors by optimizing vector yields at harvest and downstream purification steps; and 3. To manufacture, purify, release, and monitor the stability of trivalent, clinical-grade rAd26 and rAd5HVR48 vaccines expressing the optimal HIV Gag-Nef, Pol, and Env antigens under GMP conditions.