FY 2013 SBIR-I Contract for Automated Instrument to Clean Microtiter Plates

i）第1阶段的目标是开发和验证原型组件，这些组件可以很容易地集成，以创建一个旨在满足NCATS合同RFP规定的规范的仪器。 ii）通常用于高通量筛选的B1-微量滴定板包括96孔（8x 12）、384孔（16 x24）或1536孔（32 x48）板。接骨板通常具有共同的总体外部尺寸（128 mm长x 85.5 mm宽-由SBS标准定义），其中威尔斯孔以正方形网格间距为中心，该网格间距将96孔中心到中心距离（9 mm）细分为2和4倍，以生成用于 384孔板和1536孔板。孔间距的模2细分的主要实际效果是，例如设计有以9mm的线性间距布置的8或12个移液管的多组自动移液仪器也可以通过编程简单的偏移来编程以移液到384或1536孔板，所述移液管以9mm的线性间距布置，所述移液器适合于96孔板。类似地，能够同时进入384个威尔斯孔的移液头或销工具可以通过偏移编程以与1536孔板一起工作。将实现格式之间原型的互操作性。尽管板的清洁要求可能因最终用途应用而异，但理想的目标是创建一种能够清洁所有类型的板的设备，以达到同样高的标准，以允许最大的操作灵活性，而不会影响测定质量。清洁过程应尽可能保留影响试验重复性的板的关键光学和所有其他物理特征（例如，底部读取板的透明度，紧密贴合PCR板的尺寸保留等）。测试计划将确认原型组件与所有接骨板类型配合使用的有效性。 iii）B-2为了最小化液体洗涤废物，原型将利用两阶段清洁工艺，该两阶段清洁工艺使用相对简单的洗涤/漂洗工艺，用多种清洁溶液去除90-95%的板污染物，然后进行介电等离子体处理。这种设计是高质量HTS板清洁应用的有效且具有成本效益的方法。选择这种方法是因为洗涤的有效性随着浓度而下降，而等离子体清洁的有效性随着浓度的下降而增加。优化这两种方法的使用提供了显著提高的清洁水平，而与污染分子的化学性质无关。该提案没有规定关于清洗量的测量指标，但拟议的测试计划提供了广泛的测试，以确定达到清洁基准的最小量。 iv）B-3所提出的装置将板清洁过程分成两个步骤，初始洗涤/漂洗循环（如下所述），随后是单独的等离子体清洁操作。在后面的步骤中， 等离子体使板上任何残留的溶剂蒸发，并排出溶剂的气相。当板干燥时，将达到设计优化。测试计划提供了广泛的测试，以确定根据测试的应用程序达到此基准的最短时间。 v）B-4整个过程将使微板仅经受液体喷雾或等离子体气体流。本计划将仅使用此设计范例。 vi）B-5循环处理时间将少于5分钟。这正在B-2和B-3中进行测试和优化。 vii）B-6已经选择了一组严格的测定，其包括去除超过99%的生物试剂、去除超过99%的化合物以及将增强系统的市场吸引力的几种其他测定，并且还测试板的最小使用寿命，在50个循环时没有降解，以及 提供量化无菌保证水平计划的试验。所有试验的数据将纳入进度报告和I期最终报告中。 viii）B-7在我们的建议书中已经提供了初步生产的成本估算，在第1阶段，该估算将针对小批量生产进行细化。我们还将提供成本估算，以便在市场上以估计的单位数量生产该装置，这是商业化的重要基准。

i) The Phase 1 objectives are to develop and validate prototype components that may be readily integrated to create an instrument designed to meet the specifications defined by the NCATS Contract RFP. ii) B1-Microtiter plates commonly used for high throughput screening include 96 well (8x12), 384 well (16x24) or 1536 well (32x48) plates. The plates generally share common overall external dimensions (128 mm long x 85.5 mm wide - defined by the SBS Standard), with wells centered on a square grid spacing that subdivides the 96-well center-to-center distance (9mm) by a factor of 2 and 4 to generate the well center spacing for the 384 well and 1536 well plates, respectively. The major practical effect of the modulo-two subdivision of the well spacing is that multigang automatic pipetting instruments designed, for example, with 8 or 12 pipettes arranged with a linear spacing of 9mm suitable for 96-well plates, can also be programmed to pipette to 384 or 1536 well plates by programming a simple offset. Similarly, a pipetting head or pin tool able to simultaneous access 384 wells can be programmed by offset to work with a 1536-well plate. Interoperability of the prototype between the formats will be achieved. Although the cleaning requirements for plates may differ depending on end use application, the ideal objective is to create a device that is capable of cleaning all varieties of plate types to an equally high standard to allow maximum operational flexibility without risking assay quality. The cleaning process should, as far as possible, preserve key optical and all other physical features of the plate that influence assay repeatability (e.g. transparency for bottom-read plates, dimensional preservation for close-fitting PCR plates,etc.) The test plan will confirm the ultility of the prototype components to work with all plate types. iii) B-2 In order to minimize liquid wash waste the prototype will utilize a two-stage cleaning process that uses a relatively simple wash/rinse process with multiple cleaning solutions to remove 90-95% of the plate contaminants, followed by a dielectric plasma treatment. This design is an efficient and cost-effective approach for high quality HTS plate cleaning applications. This approach is selected because the effectiveness of washing declines with concentration and plasma cleaning increases in effectiveness with declining concentration. 0ptimizing the use of the two processes provides a dramatically enhanced level of cleaning irrespectiveof the chemical nature of the contaminating molecules. The proposal did not specify a metric to be measured regarding wash volumes but the testing plan proposed provides extensive testing to determine minimum volumes that will achieve the benchmark for cleaning. iv) B-3 The proposed device separates the plate cleaning process into two steps, an initial wash/rinse cycle (described below), followed by a separate plasma cleaning operation. During the later step, the energy in the plasma vaporizes any remaining solvent on the plate and the gas phase of the solvent is vented. Design optimization will be reached when the plates are dry. The testing plan provides extensive testing to determine the minimum time to achieve this benchmark based on the applications tested. v) B-4 The entire process will subject the microplates to only liquid spray or a stream of plasma gas. The plan will only use this design paradigm. vi) B-5 Cycle process time will be less than 5 minutes. This is being tested and optimized in B-2 and B-3. vii) B-6 A rigorous set of assays have been selected that include the removal to over 99% of a biological reagent, removal to over 99% of a chemical compound and several other assays that will enhance the market appeal of the system and also testing plates for a minimum useful life with no degradation at 50 cycles and tests that provide a plan to quantify a Sterility Assurance Level. Data from all testing will be included in progress reports and the Phase 1 Final Report. viii) B-7 A cost estimate has been provided for preliminary production in our proposal and during Phase 1 that estimate will be refined for low volume production. We will also provide a cost estimate to produce the unit at estimated unit quantities once on the market, which is an important benchmark for commercialization.