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ViraTag: A Scalable Purification Technology for Adeno-Associated Virus Gene Therapy Vectors

ViraTag：腺相关病毒基因治疗载体的可扩展纯化技术

基本信息

批准号：
10156608
负责人：
Kelli Michelle Luginbuhl
金额：
$ 22.49万
依托单位：
ISOLERE BIO, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-05 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10156608
关键词：
Address Affinity Affinity Chromatography Beds Behavior Benchmarking Binding Biological Products Biomanufacturing Biotechnology Buffers Capsid Capsid Proteins Cell Culture Techniques Cells Centrifugation Chromatography Clinical Collection Contracts Culture Media Custom DNA Dependovirus Development Disease Documentation Dose Elastin Engineering Environment Feedback Fermentation Filtration Gene Transduction Agent Gold Harvest Industry Laboratories Left Liquid substance Manufacturer Name Methodology Methods Peptides Performance Phage Display Pharmacologic Substance Phase Phase Transition Price Procedures Process Production Proteins Protocols documentation Reagent Recombinant Fusion Proteins Research Research Personnel Safety Scientist Serotyping Small Business Innovation Research Grant Sodium Chloride Stimulus Technology Training Treatment Efficacy Ultracentrifugation Universities Validation Viral Viral Vector Virion Virus Water aqueous bioprocess cost cost effective design environmental change flasks gene therapy innovation instrumentation novel nucleic acid-based therapeutics particle preservation product development scale up success therapeutic target vector virus culture

项目摘要

PROJECT SUMMARY Gene therapies remain promising candidates for a broad range of intractable diseases, however their complexity renders traditional biopharmaceutical manufacturing procedures inefficient, costly, and impractical. Given that a single dose of an AAV-delivered gene therapy may require purification of virus from 60 L of culture media or more, there is an urgent and unmet need to streamline the process with a scalable, high throughput, and cost- effective solution. While ultracentrifugation and chromatography are the most common methods for AAV purification, there is currently no “gold standard” method in commercial production. These limitations of current methods have motivated Isolere Bio, Inc. to develop an innovative purification platform for viruses: ViraTag™. ViraTag™ involves two sequential steps. First, the viruses are affinity captured by a custom protein reagent and the user triggers a liquid-liquid phase transition by a simple environmental change, such as the addition of salt or heat. The virus particles are sequestered into water-immiscible droplets, with host cell proteins and other cellular contaminants left behind in the aqueous phase. Second, once contaminants have been washed away, the AAV are extracted from the droplets by lowering pH, which releases them from the capture reagent. ViraTag™ addresses key requirements for an ideal purification process, including: 1) linear scalability to accommodate 103 to 105 L of input media; 2) mild elution conditions to preserve vector function and therapeutic efficacy; 3) validated compatibility with automated downstream purification instrumentation and workflows used in biomanufacturing, and 4) the potential to isolate capsids containing therapeutic nucleic acid payloads away from empty capsids. During Phase I of this SBIR Fast track proposal, we will engineer a new ViraTag™ reagent capable of affinity capture and phase separation of AAV particles, and evaluate its efficiency in AAV purification from cell culture media. We will also evaluate the ability of this new affinity capture reagent to distinguish “full” versus “empty” capsids, an urgently needed capability currently only enabled by ultracentrifugation methods that cannot be scaled up. Phase II will focus on scale-up of ViraTag™ manufacturing, by developing workflows and product form factors for various application scales, and benchmarking the purity of final products against current methods. Phase II will also include external validation of the process by laboratories representative of the product’s target customers, ranging from bench-scale academic research to large-scale biopharma.

项目总结