Development of an Advanced Screening Platform

描述（由申请人提供）：筛选仍然是发现新酶、蛋白质疗法和小分子药物的主要瓶颈。许多重要的高价值生物活性物质尚未被发现。该项目旨在克服传统筛选方法的功能和通量限制，展示百万孔筛选平台从复杂环境基因文库中发现稀有生物活性物质的可行性。Diversa的10万孔GigaMatrixTM平台已成功用于通过表达筛选发现新型酶。我们现在建议在这种筛选方法上取得重大进展。通过使用百万孔板，我们的目标是使用标准液相荧光测定每天筛选10亿个克隆的通量。这种创新的必要性来自于筛选复杂的、多生物的（10的7次方到10的10次方）DNA文库的需求，这些文库既来自原始环境样本，也来自实验室进化技术。此外，这一发展不仅可以发现罕见的新酶，还可以发现抗体和蛋白质治疗方案，这些方案需要快速筛选10的8次方到10的9次方的抗体和蛋白质变体库。从筛选中获得的生物活性分子和GigaMatrix百万孔技术本身都有望商业化，每一种技术都对新药物和治疗方法的合成做出了重大贡献。

DESCRIPTION (provided by applicant): Screening remains a major bottleneck in the discovery of new enzymes, protein therapeutics, and small molecule drugs. Many important high-value bioactives have yet to be discovered. This project is aimed at overcoming functional and throughput limitations of conventional screening methodologies by demonstrating the feasibility of a million-well screening platform for the discovery of rare bioactives from complex environmental gene libraries. Diversa's 100,000-well GigaMatrixTM platform has been successfully utilized to discover novel enzymes by expression screening. We now propose to make a significant advance in this approach to screening. By using million-well plates, we are aiming at screening throughputs of one billion clones per day using standard liquid-phase fluorescent assays. The necessity for this innovation derives from the requirement to screen complex, multi-organism (10 to the power of 7 to 10 to the power of 10) DNA libraries generated both from primary environmental samples and from laboratory evolution techniques. Furthermore, this development enables not only the discovery of rare novel enzymes but also antibody and protein therapeutic programs that require rapid screening of libraries with 10 to the power of 8 to 10 to the power of 9 antibody and protein variants. Commercialization of both bioactive molecules derived from screening and the GigaMatrix million-well technology itself is envisioned, each representing a significant contribution to the synthesis of new pharmaceuticals and therapeutics.