SBIR Phase I: New approach for producing increased quantities of human antibodies for research, medical, and biodefense applications

这个小型企业创新研究(SBIR)项目的广泛影响/商业潜力将满足抗体市场对快速和廉价生产抗体的需求。抗体是生命科学中使用最多的分子工具之一，其应用跨越了生物科学和医学的各个领域。2008年，全球抗体市场估计约为400亿美元，预计到2016年将超过680亿美元。然而，新抗体的开发是一个漫长而昂贵的过程。我们的目标是开发一种系统，显著减少开发新抗体的时间和成本。建议的系统和产生的抗体可用作未来疗法的来源，用作生物防御和医学中检测系统的组件，或用作研究和诊断的亲和纯化系统。该系统将在第一阶段进行演示，生产针对肉毒杆菌神经毒素的抗体，这将对生物防御领域具有价值。这项SBIR项目旨在开发一种针对特定应用的快速产生大量抗体的系统。第一阶段的主要目标是开发一个平台，允许健壮地产生产生大量抗体的克隆，而不需要耗时和费力的基因扩增技术，如脱氢叶酸还原酶或谷氨酰胺合成酶选择系统。这将与以前开发的一种方法相结合，该方法允许同时产生多个人源化或完全人类抗体，以快速产生能够满足抗体市场每个细分市场中存在的特定需求的不同抗体集合。然而，通过将构建体直接导入次级细胞系(如中国仓鼠卵巢(CHO)和人胚胎肾(HEK))产生的克隆产生的抗体数量比传统杂交瘤低得多。产生的抗体数量不足以满足抗体市场任何细分市场的需求。为了满足抗体市场所有细分市场的需求，建议开发一种新的方法，快速有效地产生和选择免疫球蛋白产量与普通杂交瘤相当的细胞克隆。与免疫球蛋白编码结构的强健组装系统相结合，这种方法将允许快速而强健地产生适合于特定应用的抗体。

The broader impact /commercial potential of this Small Business Innovation Research (SBIR) project will address the needs of the antibody market for fast and inexpensive production of antibodies. Antibodies are among the most utilized molecular tools in the life sciences, whose applications span a variety of areas in biological science and medicine. The worldwide antibody market was estimated at approximately $40 billion in 2008, and is expected to rise above $68 billion by 2016. The development of new antibodies, however, is a lengthy and expensive process. The goal is to develop a system that significantly reduces the time to develop new antibodies as well as the cost. The proposed system and resulting antibodies may be used as a source of future therapeutics, as components of detection systems in biodefense and medicine, or as affinity purification systems for research and diagnostics. The system will be demonstrated in Phase I with the production of antibodies targeted at botulinum neurotoxin, which will be of value to the biodefense area.This SBIR project aims to develop a system for the fast generation of high quantities of antibodies targeted at specific applications. The main objective in Phase I is to develop a platform that allows robust generation of clones producing high quantities of antibodies without the need for time-consuming and labor-intensive gene amplification techniques, such as the dehydrofolate reductase or the glutamine synthase selection systems. This will be coupled with a previously developed method that allows simultaneous generation of multiple humanized or completely human antibodies for the fast generation of diverse collections of antibodies that can address specific needs existing in each segment of the antibody market. However, clones generated via direct transfection of constructs into secondary cell lines, such as Chinese Hamster Ovary (CHO) and Human Embryo Kidney (HEK), produce substantially lower amounts of antibodies than do conventional hybridomas. The amount of antibodies produced is not sufficient to address the needs in any segment of the antibody market. To address needs existing in all segments of the antibody market, it is proposed to develop a new approach for fast and efficient generation and selection of cell clones whose immunoglobulin production levels will be comparable to those of average hybridomas. In combination with the system for robust assembly of immunoglobulin-encoding constructs, this approach will allow the rapid and robust generation of antibodies adapted for use in specific applications.