Novel Approach for Fast and Economic Manufacturing of Human Antibodies

快速、经济地生产人类抗体的新方法

基本信息

批准号：
7673898
负责人：
Alexey G Zdanovsky
金额：
$ 23.13万
依托单位：
ALPHA UNIVERSE, LLC
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7673898
关键词：
Adverse reactions Ampicillin Resistance Antibodies Antibody-Producing Cells Antigens B-Lymphocytes Bacteriophages Blood Bontoxilysin Botulism Cell Separation Cells Chimera organism Cloning Cloning Vectors Communicable Diseases Complementary DNA Cytomegalovirus DNA DNA Restriction Enzymes Development Early Promoters Economics Engineering Evaluation Expression Library Genes Genome Goals Government Green Fluorescent Proteins Human Human Development Hybridomas Immunoglobulin G Immunoglobulins Individual Infection Injectable Kanamycin Knowledge LacZ Genes Lactose Length Light Location Lymphocyte Mammalian Cell Memory Messenger RNA Methods Monoclonal Antibodies Nature Organism Peptides Peripheral Persons Pharmaceutical Preparations Phase Polyadenylation Polymerase Chain Reaction Population Price Procedures Process Production Property Protective Agents Protocols documentation Public Health Recovery Regulatory Element Replication Origin Research Research Design Reverse Transcription Risk Scheme Secure Serotyping Signal Transduction Simian virus 40 Site Steam Suggestion System Techniques Technology Testing Time Toxic effect Toxin Vaccination Vaccines Variant Work beta-Galactosidase biothreat botulinum chimeric antibody cost design endodeoxyribonuclease SceI expression vector interest novel novel strategies pathogen polyclonal antibody promoter public health relevance rapid detection research study tool vector

项目摘要

DESCRIPTION (provided by applicant): A growing number of potential native and engineered biothreat agents effectively limit the proactive protection of public health through vaccination. An alternative approach relies on the use of corresponding protective antibodies that are pre-made and are used as injectable drugs in reactive rather than proactive mode. The implementation of this approach, however, is complicated by the fact that administration of non-human antibodies can trigger development of adverse reactions. Securing of human antibodies in sufficient quantities so far has been a challenging task. Also, the traditional approach of producing antibodies depends on the existence of corresponding vaccines and cannot be used for expedient development of protective agents against newly emerging infections. To overcome these hurdles we propose to develop a novel system for assembly and production of human/human chimeric antibodies. Such antibodies can be generated at a fraction of the cost of raising polyclonal antibodies and in substantially shorter times than traditional monoclonal antibodies while still remaining human in nature. In general, our system will be much safer to operate with than hybridoma technology. In addition, it can be used for development of protective entities at the first signs of emergence of new threat agents. Phase I efforts will test the feasibility of our approach by developing human/human chimeras that will recognize botulinum neurotoxin serotype A. To assemble such chimeras, we will introduce a proprietary cloning scheme that reduces the risk of modifying native antigen-recognizing sequences and substantially simplifies the process of assembly of immunoglobulin-encoding sequences. This will make the system suitable for high-throughput format applications. Regulatory elements incorporated into cloning vectors will enable rapid detection of cells producing antibodies of interest and selection of host cells for industrial production, thus increasing the efficiency of the process and decreasing price of the final product. During Phase II, the developed cloning approach and expression vectors will be used to generate IgG variants of chimeric antibodies that will be able to neutralize toxic effects of botulinum neurotoxin serotypes A and B and will be suitable for use as anti-botulinum drugs. The developed enabling technology will be suitable for generating protective antibodies against other biothreat agents. PUBLIC HEALTH RELEVANCE: The current project will provide a new way for development of human anti-pathogen antibodies and opens new opportunities for passive vaccination as a way of protecting humans against infectious diseases and toxins.

描述（由申请人提供）：越来越多的潜在天然和工程生物威胁剂有效限制了通过疫苗接种对公众健康的主动保护。另一种方法依赖于使用预先制备的相应保护性抗体，并以反应性而非主动模式用作注射药物。然而，由于施用非人类抗体可能引发不良反应，因此该方法的实施变得复杂。迄今为止，获得足够数量的人类抗体一直是一项具有挑战性的任务。此外，传统的抗体生产方法依赖于相应疫苗的存在，无法用于快速开发针对新出现的感染的保护剂。为了克服这些障碍，我们建议开发一种用于组装和生产人/人嵌合抗体的新系统。与传统的单克隆抗体相比，此类抗体的生产成本仅为多克隆抗体的一小部分，且所需时间大大缩短，同时仍保持人类的本质。一般来说，我们的系统比杂交瘤技术操作起来更安全。此外，它还可用于在新威胁代理出现的最初迹象时开发保护实体。第一阶段的工作将通过开发可识别肉毒杆菌神经毒素 A 血清型的人/人嵌合体来测试我们方法的可行性。为了组装此类嵌合体，我们将引入一种专有的克隆方案，该方案可降低修改天然抗原识别序列的风险并大大简化免疫球蛋白编码序列的组装过程。这将使该系统适合高通量格式应用。整合到克隆载体中的调控元件将能够快速检测产生感兴趣抗体的细胞并选择用于工业生产的宿主细胞，从而提高过程效率并降低最终产品的价格。在第二阶段，开发的克隆方法和表达载体将用于产生嵌合抗体的IgG变体，这些变体将能够中和肉毒杆菌神经毒素血清型A和B的毒性作用，并且适合用作抗肉毒杆菌药物。开发的使能技术将适用于生成针对其他生物威胁剂的保护性抗体。公共卫生相关性：当前的项目将为人类抗病原体抗体的开发提供一种新方法，并为被动疫苗接种开辟新的机会，作为保护人类免受传染病和毒素的一种方式。