Multiplexed screening of protein expression libraries

描述（由申请方提供）：在本项目的I期，Trellis Bioscience，Inc.已经验证了一种专有的筛选技术，用于对来自单个活细胞的分泌蛋白进行多重表征，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，实现了特异性（质量）阈值，明显高于其他抗体筛选技术的可行性。鉴定的克隆太罕见，无法通过缺乏网格方法的极端小型化（100，000倍减少）的常规方法进行回收。目前的II期资金申请是为了将该技术从鼠杂交瘤扩展到两个独特的多样性来源：重组抗体和重组小肽。每一种都解决了鼠杂交瘤不能满足的市场需求。成功地延伸到重组抗体将允许针对传染病的人源抗体的回收，将疫苗接种的益处延伸到大规模接种活动不切实际或不合适的疾病。肽对于靶向不适合抗体治疗的细胞内药物靶标特别感兴趣。扩展到这些用途需要大幅度增加筛选系统的吞吐量。因此，目标1侧重于增加分析的多路复用能力和提高图像分析系统的效率和通量，而目标2和3侧重于调整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治疗将允许有效开发药物来治疗目前不适合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.