Design of synthetic recombinant antibody devices for applications in molecular targeted imaging
用于分子靶向成像应用的合成重组抗体装置的设计
基本信息
- 批准号:RGPIN-2020-06194
- 负责人:
- 金额:$ 2.11万
- 依托单位:
- 依托单位国家:加拿大
- 项目类别:Discovery Grants Program - Individual
- 财政年份:2022
- 资助国家:加拿大
- 起止时间:2022-01-01 至 2023-12-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Antibodies are large in size and possess a crystallizable fragment (Fc) domain, resulting in low tissue penetration, long in vivo half lives, and expensive and sometimes difficult production methods, which are often not suitable for many applications. Recombinant antibody (rAb) fragments have been engineered to overcome these limitations by controlling rAb size and configurations, binding affinities and specificities, Fc domain properties, and by fusing rAbs to other proteins or tags that facilitate detection and purification. These rAb fragments often require sophisticated antibody architectures not present in nature, which can result in low expression, poor solubility and aggregation, and toxicity. Thus, many rAb fragments require expensive and complex manufacturing that must be optimized for each rAb fragment. We developed a novel approach to construct rAb-like proteins using a "bottom up" synthetic strategy with discrete well behaved antibody domain or "parts". To do this, we express and post translationally assemble antibody constant and variable domain parts in different combinations using a protein ligase to create rAb like devices from scratch with desired properties that are based on the summed properties of the individual parts. We use the SpyTag/SpyCatcher ligase system to post-translationally assemble antibody parts and other protein/peptide domains in different combinations, some of which are not possible to generate in nature or using genetic engineering approaches. The long term objective of our research program is to develop an efficient, cost effective strategy for assembling rAb-like devices from discrete antibody domain parts for applications in basic science and engineering, diagnostics, and therapy. To achieve this, we are pursuing our short-term objective of designing, constructing, purifying, and validating rAb-like devices for molecular targeted and pre-targeted imaging. We propose to establish a library of antibody domain parts that can be assembled post-translationally into complex devices using the SpyTag/SpyCatcher protein ligase system. To achieve this short-term objective, we have established the following aims: (i) Develop an improved phage display platform for generating variable domain parts; (ii) Establish methods to use the SpyTag/SpyCatcher system to assemble multivalent probes for molecular imaging; (iii) Improve the in vivo SpyTag/SpyCatcher ligation; and (iv) Validate multivalent rAbs for molecular targeted and pretargeted imaging. This synthetic biology approach for the modular assembly of rAbs will improve yields of rAbs that are difficult to express, provide rapid methods to produce rAbs, and allow antibody domains to be connected in ways that cannot be obtained by using genetic engineering approaches. Using easy to produce and well characterized antibody parts to construct rAb devices will reduce development costs and time, and improve the safety of rAbs used in imaging and therapy.
抗体尺寸大并且具有可结晶片段(Fc)结构域,导致低组织渗透性、长体内半衰期以及昂贵且有时困难的生产方法,其通常不适合于许多应用。重组抗体(rAb)片段已经被工程化以通过控制rAb大小和构型、结合亲和力和特异性、Fc结构域特性以及通过将rAb融合到促进检测和纯化的其他蛋白质或标签来克服这些限制。这些rAb片段通常需要自然界中不存在的复杂抗体结构,这可能导致低表达、溶解性和聚集性差以及毒性。因此,许多rAb片段需要昂贵且复杂的制造,其必须针对每个rAb片段进行优化。我们开发了一种新的方法来构建rAb样蛋白,使用“自下而上”的合成策略与离散的表现良好的抗体结构域或“部分”。为了做到这一点,我们使用蛋白质连接酶以不同的组合表达和后组装抗体恒定和可变结构域部分,以从头开始创建具有基于各个部分的总和性质的所需性质的rAb样装置。我们使用SpyTag/SpyCatcher连接酶系统以不同的组合后组装抗体部分和其他蛋白质/肽结构域,其中一些是不可能在自然界中或使用基因工程方法产生的。我们研究计划的长期目标是开发一种有效的,具有成本效益的策略,用于从离散的抗体结构域部分组装rAb样装置,用于基础科学和工程,诊断和治疗。为了实现这一目标,我们正在追求我们的短期目标,即设计、构建、纯化和验证用于分子靶向和预靶向成像的rAb样装置。我们建议建立一个库的抗体结构域部分,可以组装成复杂的设备使用SpyTag/SpyCatcher蛋白质连接酶系统的post-acquisitionally。为了实现这一短期目标,我们已经建立了以下目标:(i)开发用于产生可变结构域部分的改进的噬菌体展示平台;(ii)建立使用SpyTag/SpyCatcher系统组装用于分子成像的多价探针的方法;(iii)改进体内SpyTag/SpyCatcher连接;以及(iv)制备用于分子靶向和预靶向成像的多价rAb。这种用于rAb模块化组装的合成生物学方法将提高难以表达的rAb的产量,提供快速生产rAb的方法,并允许抗体结构域以无法通过使用基因工程方法获得的方式连接。使用易于生产和良好表征的抗体部分来构建rAb装置将降低开发成本和时间,并提高用于成像和治疗的rAb的安全性。
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)
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Geyer, Clarence其他文献
Geyer, Clarence的其他文献
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{{ truncateString('Geyer, Clarence', 18)}}的其他基金
Design of synthetic recombinant antibody devices for applications in molecular targeted imaging
用于分子靶向成像应用的合成重组抗体装置的设计
- 批准号:
RGPIN-2020-06194 - 财政年份:2021
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Design of synthetic recombinant antibody devices for applications in molecular targeted imaging
用于分子靶向成像应用的合成重组抗体装置的设计
- 批准号:
RGPIN-2020-06194 - 财政年份:2020
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Intracellular cyclic reagents for validating protein function
用于验证蛋白质功能的细胞内循环试剂
- 批准号:
249793-2008 - 财政年份:2009
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Intracellular cyclic reagents for validating protein function
用于验证蛋白质功能的细胞内循环试剂
- 批准号:
249793-2008 - 财政年份:2008
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Peptide aptamer-based protein-detecting arrays
基于肽适体的蛋白质检测阵列
- 批准号:
249793-2003 - 财政年份:2007
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Peptide aptamer-based protein-detecting arrays
基于肽适体的蛋白质检测阵列
- 批准号:
249793-2003 - 财政年份:2005
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Peptide aptamer-based protein-detecting arrays
基于肽适体的蛋白质检测阵列
- 批准号:
249793-2003 - 财政年份:2004
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
Peptide aptamer-based protein-detecting arrays
基于肽适体的蛋白质检测阵列
- 批准号:
249793-2003 - 财政年份:2003
- 资助金额:
$ 2.11万 - 项目类别:
Discovery Grants Program - Individual
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Design of synthetic recombinant antibody devices for applications in molecular targeted imaging
用于分子靶向成像应用的合成重组抗体装置的设计
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