Multiplexed screening of protein expression libraries

描述（由申请人提供）：在该项目的第一阶段，Trellis Bioscience, Inc.已经验证了一种专利筛选技术，用于从单个活细胞中多重表征分泌蛋白，从而能够检测和繁殖罕见的有利细胞。该技术应用于小鼠脾细胞和杂交瘤，达到特异性（质量）阈值，明显高于其他抗体筛选技术的可行性。已鉴定的克隆过于罕见，无法通过传统方法恢复，缺乏Trellis方法的极端小型化（减少100,000倍）。目前的第二阶段资金申请是将该技术从小鼠杂杂瘤扩展到两种不同的多样性来源：重组抗体和重组小肽。每一种都满足了鼠杂交瘤无法满足的市场需求。成功地推广到重组抗体将使防治传染病的人源抗体得以恢复，将疫苗接种的好处扩大到大规模接种运动不切实际或不适当的疾病。肽是特别感兴趣的针对细胞内的药物靶标不适合抗体治疗。扩展这些用途需要大幅增加筛选系统的吞吐量。因此，目标1侧重于增加检测的多路复用能力，提高图像分析系统的效率和吞吐量，而目标2和目标3侧重于适应该技术并展示其在两个感兴趣领域的能力。Trellis Bioscience正在开发一种技术，可以减少发现抗体疗法所需的时间和成本，抗体疗法现在是制药行业增长最快的部分。这类药物被用于治疗癌症和炎症性疾病，以及传染病。将该技术进一步扩展到肽疗法将允许有效地开发药物来治疗目前无法采用基于抗体的方法的疾病。

DESCRIPTION (provided by applicant): In Phase I of this project, Trellis Bioscience, Inc. has validated a proprietary screening technology for multiplexed characterization of the secreted protein from individual live cells, enabling detection and propagation of rare favorable cells. The technology was applied to murine splenocytes and hybridomas, achieving a specificity (quality) threshold that is markedly higher than is feasible for other antibody screening technologies. The identified clones were too rare for recovery by conventional means lacking the extreme miniaturization of the Trellis approach (100,000 fold reduction). The present Phase II request for funding is to enable extension of the technology from murine hybridomas to two distinctive diversity sources: recombinant antibodies, and recombinant small peptides. Each addresses market needs not satisfied by murine hybridomas. Successful extension to recombinant antibodies will allow the recovery of human derived antibodies against infectious diseases, extending the benefits of vaccination to diseases for which a mass innoculation campaign is impractical or inappropriate. Peptides are of particular interest for targeting intracellular drug targets not amenable to antibody treatment. Extension to these uses requires a substantive increase in throughput of the screening system. Accordingly, Aim 1 is focused on increasing the multiplexing capacity of the assay and improving the efficiency and throughput of the image analysis system, while Aims 2 and 3 are focused on adapting the technology and demonstrating its capabilities in the two fields of interest. Trellis Bioscience is developing a technology that decreases the time and cost required to discover antibody therapeutics, which now represent the fastest growing segment of the pharmaceutical industry. Drugs in this class are being used to treat cancer and inflammatory diseases, as well as infectious disease. A further extension of the technology to peptide therapeutics will allow the efficient development of drugs to treat diseases that are not currently amenable to antibody based approaches.