The development of a microscopy-based cell-array toxicity assay for quantifying C

描述（由申请人提供）：活细胞筛选代表了新药和生物制剂开发的重大范式转变。尽管为此目的取得了许多自动化和数据分析方面的进步，但许多挑战仍然阻碍了这种方法的实施。作为一组实例，用于评价抗体药物功效的最常见细胞毒性测定，补体依赖性细胞毒性（CDC）和抗体依赖性细胞毒性（ADCC）测定，仅关注终点结果。因此，它们几乎不能提供关于异质性肿瘤群体中哪些细胞被杀死或耐药的信息。此外，目前提供的检测产品都不适合使用高含量成像来研究细胞毒性的发生和速率，尽管这种方法可以提供丰富的数据。Adheren的新细胞阵列毒性 (CAT)通过最小化假阳性、提供更早的细胞毒性数据和允许真实的可视化细胞死亡，测定平台有望推进该领域。这些产品基于核心的DNA细胞附着平台，该平台允许对非常广泛的细胞类型进行强大和可编程的细胞结合。粘附方法允许活细胞和杀死的细胞都保留在微阵列上，提供改进的细胞毒性统计。 该技术还允许洗涤细胞微阵列而没有细胞损失，并且它能够使用荧光成像随时间直接可视化细胞毒性。这可以为阐明基于补体和效应细胞的细胞毒性机制提供前所未有的实时细胞特异性数据水平。我们建议通过验证它们对广泛的细胞类型和生物测定的有效性来扩大和进一步开发这些基于细胞阵列的ADCC和CDC显微镜测定。此外，试验平台将从目前使用的玻璃底96孔板扩展到包括低成本工业标准聚苯乙烯96孔板。

DESCRIPTION (provided by applicant): Live cell screening represents a major paradigm shift for the development of new pharmaceuticals and biologics. Despite the many automation and data analysis advances for this purpose, however, many challenges still hinder the implementation of this approach. As one set of examples, the most common cell cytotoxicity assays used to evaluate the efficacy of an antibody drug, the complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) assays, only focus on end-point results. As a result, they yield little information about which cells are killed or resitant in a heterogeneous tumor population. In addition, none of the assay products offered today are compatible with the use of high content imaging to study the onset and rate of cytotoxicity, despite the rich quantity of data that this approach can provide. Adheren's new cell-array toxicity (CAT) assay platforms promise to advance this field by minimizing false positives, providing earlier cytotoxicity data, and allowing the visualization of cell death in real time. These producs are based on a core DNA-based cell attachment platform, which allows strong and programmable cell binding for a very wide range of cell types. The adhesion method allows both live and killed cells to be retained on the microarray, providing improved cytotoxicity statistics. This technology also allows the cell microarray to be washed without cell loss, and it enables the direct visualization of cell cytotoxicity over time using fluorescence imaging. This can providing unprecedented levels of real-time cell-specific data for the elucidation of complement- and effector cell-based cytotoxicity mechanisms. We propose to scale up and further develop these cell array-based ADCC and CDC microscopy assays by validating their effectiveness for a wide range of cell types and biological assays. In addition, the assay platform will be extended from currently the used glass bottom 96 well plates to include low-cost industry standard polystyrene 96 well plates.