Novel Materials for Viral Purification
用于病毒纯化的新型材料
基本信息
- 批准号:9408588
- 负责人:
- 金额:$ 22.42万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2017
- 资助国家:美国
- 起止时间:2017-09-01 至 2019-04-30
- 项目状态:已结题
- 来源:
- 关键词:AdenovirusesAffinityAreaAvidityBindingCancer VaccinesCapsid ProteinsCell Culture TechniquesCell TherapyCellsChromatographyColumn ChromatographyCommunicable DiseasesConcatenated DNACouplingDNADNA LibraryDependovirusDevelopmentDiseaseDivalent CationsExcisionExhibitsFiltrationFutureGene Transduction AgentGoalsImmobilizationIndividualLabelLibrariesMagnetismMalignant NeoplasmsMembraneMethodsModelingNucleic AcidsOligonucleotidesOncolyticPreparationProcessProductionProteinsProtocols documentationReagentRecoveryResidual stateSepharoseSingle-Stranded DNASourceSpecificityStreamStructureSubfamily lentivirinaeTechniquesTechnologyTestingViralViral VectorVirusViscosityWorkbasebioprocesscancer therapycommercial applicationcostfeedinggene therapyinterestmagnetic beadsmagnetite ferrosoferric oxidenanoparticulatenovelnucleaseoperationparticletoolvector
项目摘要
Abstract
Viruses have significant commercial application as vectors for gene and cell therapy, as well as vaccines for
cancer treatment and infectious diseases. The purification of commercial viruses is a major area of bioprocess
development. Current methods are mostly based on filtration and chromatography, both of which scale poorly
and require clean feed streams to perform well. Viral particles are difficult to recover from culture supernatants
or cell lysates because they are similar in size to cell debris. A large amount of expensive, high-quality
nuclease (Benzonase) is typically needed to reduce the viscosity of the lysate sufficiently for efficient
membrane and column operations. Affinity based vector purification has been developed for only one type of
vector (some strains of adeno-associated virus), and is also very expensive. In summary, virus purification is
challenging and significant amounts of valuable product are lost in inefficient filters.
We have developed a novel DNA-based avidity reagent, termed DeNAno, that can provide a significant
advance in the development and manufacturing of commercial viruses, with applicability to a variety of viral
vectors. DeNAno uses massive avidity rather than affinity to capture biologic targets such as viruses. DeNAno
particles are composed of a single-stranded DNA concatemer, which contains several hundred copies of a
template oligonucleotide. The goal of this project is to develop DeNAno particles that bind to a target virus, and
then use the DeNAno as a capture reagent for virus purification. DeNAno libraries are produced by rolling
circle replication of a circular oligonucleotide template containing a random sequence, and specific viral
binders are recovered by a biopanning process. DeNAno particles can be released from their targets by
disrupting their secondary structure through divalent cation removal; they can also be labeled with nano-
particulate magnetite or coated onto micron-sized magnetite to allow their use as magnetic capture reagents.
We have already performed a selection on adenovirus-coated beads and obtained a pool of DeNAno particles
that bind the target virus. The specific aims of this project are to: 1) characterize DeNAno particles that bind to
the model virus (adenovirus) and 2) demonstrate their use as a purification tool by attaching them to magnetite
so that they can be used in magnetic capture. The virus will be released following capture by divalent cation
removal and the magnetic DeNAno cleared from the viral preparation, again by magnet. We will assess the
capture efficiency, virus yield and infectivity, residual DeNAno contamination, and the presence of host cell
proteins in the final preparation. If successful, this project will demonstrate the potential of the DeNAno
technology to streamline virus production and pave the way for additional DeNAno-based applications
targeting other commercially important viruses.
抽象的
病毒作为基因和细胞治疗的载体以及疫苗具有重要的商业应用
癌症治疗和传染病。商业病毒的纯化是生物过程的一个主要领域
发展。目前的方法主要基于过滤和色谱法,这两种方法的扩展性都很差
并且需要清洁的进料流才能良好运行。病毒颗粒很难从培养上清液中回收
或细胞裂解物,因为它们的大小与细胞碎片相似。量大价高,品质优良
通常需要核酸酶(Benzonase)来充分降低裂解物的粘度,以有效地
膜和柱操作。基于亲和力的载体纯化仅针对一种类型的
载体(某些腺相关病毒株),而且也非常昂贵。综上所述,病毒纯化是
具有挑战性的且大量有价值的产品在低效的过滤器中丢失。
我们开发了一种新型的基于 DNA 的亲和力试剂,称为 DeNAno,它可以提供显着的亲和力。
商业病毒的开发和制造取得进展,适用于多种病毒
向量。 DeNAno 使用大量的亲和力而不是亲和力来捕获病毒等生物目标。德纳诺
颗粒由单链DNA串联体组成,其中包含数百个拷贝
模板寡核苷酸。该项目的目标是开发与目标病毒结合的 DeNAno 颗粒,以及
然后使用 DeNAno 作为病毒纯化的捕获试剂。 DeNAno 文库是通过滚动生成的
含有随机序列和特定病毒的环状寡核苷酸模板的环状复制
通过生物淘选过程回收粘合剂。 DeNAno 粒子可以通过以下方式从其目标释放:
通过去除二价阳离子破坏其二级结构;它们也可以用纳米标记
颗粒状磁铁矿或涂覆在微米尺寸的磁铁矿上,以使其用作磁性捕获试剂。
我们已经对腺病毒包被的珠子进行了选择,并获得了 DeNAno 颗粒池
结合目标病毒。该项目的具体目标是:1)表征结合到的 DeNAno 粒子
模型病毒(腺病毒)和 2) 通过将它们附着在磁铁矿上来展示它们作为纯化工具的用途
以便它们可以用于磁捕获。病毒将被二价阳离子捕获后释放
去除并再次通过磁铁从病毒制剂中清除磁性 DeNAno。我们将评估
捕获效率、病毒产量和感染性、残留的 DeNAno 污染以及宿主细胞的存在
最终制剂中的蛋白质。如果成功,该项目将展示 DeNAno 的潜力
简化病毒生产并为其他基于 DeNAno 的应用铺平道路的技术
针对其他商业上重要的病毒。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
BRADLEY T MESSMER其他文献
BRADLEY T MESSMER的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('BRADLEY T MESSMER', 18)}}的其他基金
Personalized precision dosing of anti-TNF biologic therapies
抗 TNF 生物疗法的个性化精确剂量
- 批准号:
9888300 - 财政年份:2018
- 资助金额:
$ 22.42万 - 项目类别:
Personalized precision dosing of biologic therapies in oncology
肿瘤学生物疗法的个性化精确剂量
- 批准号:
9984616 - 财政年份:2017
- 资助金额:
$ 22.42万 - 项目类别:
Activin A antagonist for treatment of cancer-associated cachexia
激活素 A 拮抗剂用于治疗癌症相关恶病质
- 批准号:
9046615 - 财政年份:2015
- 资助金额:
$ 22.42万 - 项目类别:
Cleavage Coupled Lateral Flow Immunoassay for Rapid Endotoxin Testing
用于快速内毒素检测的裂解偶联侧向层析免疫分析
- 批准号:
8905383 - 财政年份:2015
- 资助金额:
$ 22.42万 - 项目类别:
Multiplexed DeNAno Protein Assay and Quantitation: Sequencing Based Proteomics
多重 DeNAno 蛋白质测定和定量:基于测序的蛋白质组学
- 批准号:
8855369 - 财政年份:2015
- 资助金额:
$ 22.42万 - 项目类别:
Lateral Flow Immunoassay for Therapeutic Monoclonal Antibody Quality Assurance
用于治疗性单克隆抗体质量保证的侧流免疫分析
- 批准号:
8648070 - 财政年份:2014
- 资助金额:
$ 22.42万 - 项目类别:
Molecular Evolution of Multifunctional DNA Nanoparticles
多功能DNA纳米颗粒的分子进化
- 批准号:
8293019 - 财政年份:2011
- 资助金额:
$ 22.42万 - 项目类别:
Molecular Evolution of Multifunctional DNA Nanoparticles
多功能DNA纳米颗粒的分子进化
- 批准号:
8472338 - 财政年份:2011
- 资助金额:
$ 22.42万 - 项目类别:
Molecular Evolution of Multifunctional DNA Nanoparticles
多功能DNA纳米颗粒的分子进化
- 批准号:
8035223 - 财政年份:2011
- 资助金额:
$ 22.42万 - 项目类别:
相似海外基金
Construction of affinity sensors using high-speed oscillation of nanomaterials
利用纳米材料高速振荡构建亲和传感器
- 批准号:
23H01982 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Affinity evaluation for development of polymer nanocomposites with high thermal conductivity and interfacial molecular design
高导热率聚合物纳米复合材料开发和界面分子设计的亲和力评估
- 批准号:
23KJ0116 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Grant-in-Aid for JSPS Fellows
Platform for the High Throughput Generation and Validation of Affinity Reagents
用于高通量生成和亲和试剂验证的平台
- 批准号:
10598276 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Development of High-Affinity and Selective Ligands as a Pharmacological Tool for the Dopamine D4 Receptor (D4R) Subtype Variants
开发高亲和力和选择性配体作为多巴胺 D4 受体 (D4R) 亚型变体的药理学工具
- 批准号:
10682794 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Collaborative Research: DESIGN: Co-creation of affinity groups to facilitate diverse & inclusive ornithological societies
合作研究:设计:共同创建亲和团体以促进多元化
- 批准号:
2233343 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Standard Grant
Collaborative Research: DESIGN: Co-creation of affinity groups to facilitate diverse & inclusive ornithological societies
合作研究:设计:共同创建亲和团体以促进多元化
- 批准号:
2233342 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Standard Grant
Molecular mechanisms underlying high-affinity and isotype switched antibody responses
高亲和力和同种型转换抗体反应的分子机制
- 批准号:
479363 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Operating Grants
Deconstructed T cell antigen recognition: Separation of affinity from bond lifetime
解构 T 细胞抗原识别:亲和力与键寿命的分离
- 批准号:
10681989 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
CAREER: Engineered Affinity-Based Biomaterials for Harnessing the Stem Cell Secretome
职业:基于亲和力的工程生物材料用于利用干细胞分泌组
- 批准号:
2237240 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Continuing Grant
ADVANCE Partnership: Leveraging Intersectionality and Engineering Affinity groups in Industrial Engineering and Operations Research (LINEAGE)
ADVANCE 合作伙伴关系:利用工业工程和运筹学 (LINEAGE) 领域的交叉性和工程亲和力团体
- 批准号:
2305592 - 财政年份:2023
- 资助金额:
$ 22.42万 - 项目类别:
Continuing Grant














{{item.name}}会员




