University Hospital, Immunology Laboratory, FRE-CNRS 3312, Saint-Etienne, France.
Stem Cell Laboratory, Immunology Department, Faculty of Medicine, Yeditepe University, Istanbul, Turkey.
Front Immunol. 2020 Sep 17;11:2169. doi: 10.3389/fimmu.2020.02169. eCollection 2020.
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, 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，如基于抗体的疗法、细胞疗法（例如，基因修饰细胞、细胞因子信号调节剂和信号转导调节剂）以及其他基于生物学的干预措施，都会强烈影响免疫系统，因此需要用于研究、诊断和监测的工具。在流式细胞术中，制造商编制和验证的诊断（IVD）检测试剂盒并不能满足所有要求。因此，实验室通常依赖于修改商业上可用的检测方法，或者最常见的是开发它们以满足临床需求。然而，这两种方法都必须经过全面验证，以满足 IVD 监管要求。流式细胞术免疫表型分析是对多个参数的多参数分析，其中一些参数必须反复调整；在开发特定的抗体组合时必须考虑到这一点。在分析这些检测方法时，需要仔细调整一般规则，以满足法律和监管要求。在这里，我们描述了基于流式细胞术的检测方法的相关监管框架，并描述了将新的抗体组合引入常规工作的方法，包括开发性能规格、验证以及用于设计和分析实验的统计方法。目的是在引入内部开发的流式细胞术检测方法后提高可靠性、效率和可审核性。
