Many anticancer therapies such as antibody-based therapies, cellular therapeutics (e.g., genetically modified cells, regulators of cytokine signaling, and signal transduction), and other biologically tailored interventions strongly influence the immune system and require tools for research, diagnosis, and monitoring. In flow cytometry, in vitro diagnostic (IVD) test kits that have been compiled and validated by the manufacturer are not available for all requirements. Laboratories are therefore usually dependent on modifying commercially available assays or, most often, developing them to meet clinical needs. However, both variants must then undergo full validation to fulfill the IVD regulatory requirements. Flow cytometric immunophenotyping is a multiparametric analysis of parameters, some of which have to be repeatedly adjusted; that must be considered when developing specific antibody panels. Careful adjustments of general rules are required to meet legal and regulatory requirements in the analysis of these assays. Here, we describe the relevant regulatory framework for flow cytometry-based assays and describe methods for the introduction of new antibody combinations into routine work including development of performance specifications, validation, and statistical methodology for design and analysis of the experiments. The aim is to increase reliability, efficiency, and auditability after the introduction of in-house-developed flow cytometry assays.

许多抗癌疗法，例如基于抗体的疗法，细胞疗法（例如，基因修饰的细胞，细胞因子信号调节剂和信号转导），以及其他针对生物学的干预措施，都会强烈影响免疫系统，并需要研究，诊断和监测工具。在流式细胞仪中体外由制造商编译和验证的诊断（IVD）测试工具包并非对所有要求都可用。因此，实验室通常依赖于修改可商购的测定方法，或者最经常地是开发它们以满足临床需求。但是，这两种变体都必须经过全面验证才能满足IVD法规要求。流式细胞仪免疫表型分析是对参数的多参数分析，其中一些参数必须反复调整。开发特异性抗体组时必须考虑到这一点。在分析这些分析时，需要仔细调整一般规则，以满足法律和法规要求。这里，我们描述了基于流式细胞术的分析的相关监管框架，并介绍了将新抗体组合引入常规工作的方法，包括性能规格的开发，验证以及用于设计和分析的统计方法。目的是在引入内部开发的流式细胞仪检测后提高可靠性，效率和可审计性。