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

• 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
• 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

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地限制了通过疫苗接种对公共卫生的主动保护。另一种方法依赖于使用相应的保护性抗体，这些抗体是预先制备的，并以反应性而非主动模式用作可注射药物。然而，这种方法的实施由于施用非人抗体可能引发不良反应的发生而变得复杂。到目前为止，获得足够数量的人抗体一直是一项具有挑战性的任务。此外，产生抗体的传统方法取决于相应疫苗的存在，并且不能用于针对新出现的感染的保护剂的有利开发。为了克服这些障碍，我们提出开发一种新的系统，用于组装和生产人/人嵌合抗体。这样的抗体可以以产生多克隆抗体的成本的一小部分产生，并且比传统的单克隆抗体在实质上更短的时间内产生，同时仍然保持人的本性。总的来说，我们的系统比杂交瘤技术更安全。此外，它还可以用于在出现新的威胁物质的最初迹象时开发保护性实体。 第一阶段的工作将测试我们的方法的可行性，通过开发人类/人类嵌合体，将识别肉毒杆菌神经毒素血清型A。为了组装这样的嵌合体，我们将引入一种专有的克隆方案，该方案降低了修饰天然抗原识别序列的风险，并大大简化了免疫球蛋白编码序列的组装过程。这将使该系统适用于高通量格式应用。掺入克隆载体中的调节元件将使得能够快速检测产生感兴趣的抗体的细胞并选择用于工业生产的宿主细胞，从而提高该过程的效率并降低最终产品的价格。在II期期间，所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，所述嵌合抗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，并且将适合用作抗肉毒杆菌药物。开发的使能技术将适用于产生针对其他生物威胁剂的保护性抗体。 公共卫生相关性：目前的项目将为人类抗病原体抗体的开发提供一种新的方法，并为被动疫苗接种提供新的机会，以保护人类免受传染病和毒素的侵害。