High Precision Bioassay Using Robotics and Statistics

描述（由申请人提供）：即使经过大量的开发努力，细胞培养生物测定也被认为是费力和不精确的。这些生物测定法在生物技术工业中广泛用于测量开发中的蛋白质或疫苗产品和批量放行。不精确的检测和缓慢的检测开发导致产品开发缓慢。细胞培养生物测定的常见做法忽略了统计和监管指导，没有利用适当的随机化，也没有考虑群体效应（由多通道移液器引起）。此外，在分析中缺乏对位置效应的关注，再加上对分析数据的简单分析，损害了细胞培养生物分析的准确性和效率。我们提出了一种现代标准化的细胞培养生物测定方法，使用与实验室机器人相关的统计设计和分析，这将解决随机化、位置效应、连续稀释和分组稀释。更大的项目目标是使用统计软件和实验室机器人建立一个交钥匙细胞培养生物测定系统。本提案中描述的工作将使用1)标准化设计和分析，2)用于生物测定分析的线性或非线性混合分割或条形图模型，3)使随机化实用的实验室机器人技术的最新发展（硬件和软件），以及4)模拟连续稀释误差的新代码（在EPSCoR资助下开发，Lansky 2004）。这些方法的组合，一旦开发出来，应该将变异系数从高个位数或高得多降低到低个位数，将复制的需要减少四到十倍或更多。最重要的是，通过建立一个交钥匙机器人和分析生物测定系统，这种水平的测定性能将广泛地提供给生物技术行业以及非营利研究人员。

DESCRIPTION (provided by applicant): Cell culture bioassays are perceived as laborious and imprecise even after substantial development efforts. These bioassays are used broadly in the biotechnology industry to measure protein or vaccine products in development and for lot release. Imprecise assays and slow assay development contribute to slow product development. Common practice in cell culture bioassay ignores statistical and regulatory guidance by failing to utilize proper randomization and not accounting for group effects (caused by multi-channel pipettes). In addition, the lack of attention to location effects within the assays combined with simplistic analyses of assay data compromise the precision and efficiency of cell culture bioassays. We propose a modern standardized approach to cell culture bioassay using statistical design and analysis linked to laboratory robots, which will address randomization, location effects, serial dilution, and grouped dilution. The larger program goal is to build a turnkey cell culture bioassay system using statistical software and a laboratory robot. The work described in this proposal will use 1) standardized designs and analysis, 2) linear or nonlinear mixed split- or strip-plot models for bioassay analysis, 3) recent developments in laboratory robotics that make randomization practical (both hardware and software), and 4) new code for modeling serial dilution error (developed under EPSCoR funding, Lansky 2004). The combination of methods, once developed, should reduce the coefficient of variation from high single digits or substantially higher to low single digits, reducing the need for replication by a factor of four to ten or more. Most importantly, by building a turnkey robotic and analysis bioassay system, this level of assay performance will be made broadly available to the biotechnology industry as well as to non-profit researchers.