Yi Kaijuan, Wang Can, Lu Huili
Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240, China.
NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai Institute for Food and Drug Control, Shanghai, 201203, China.
Bioresour Bioprocess. 2025 Sep 24;12(1):103. doi: 10.1186/s40643-025-00932-2.
The assessment of biological product activity is a key aspect of quality control. Currently, in vitro assays serve as the primarily method employed by both companies and regulatory agencies to evaluate biological activity. Reporter Gene Assay (RGA) is a technique that investigates gene expression regulation and cellular signal transduction pathway activation through easily detectable reporter genes. RGA is highly dependent on drug mechanisms, offering high accuracy and precision, and has gained increasing recognition. The utilization of alternative analytical methods based on RGA have emerged as a prevailing trend, with a growing number of antibody drugs adopting corresponding RGA-based quality control approaches. Establishing stable expressing cell lines is essential to ensure the stability, reliability, and consistency of assays across diverse conditions when employing RGA techniques. CRISPR/Cas9 gene editing technology mediated site-specific gene integration allows for rapid and precise insertion of exogenous genes into specific genomic loci and enables the efficient construction of stable RGA cell lines, which would significantly propel the advancement of biological activity evaluation methods.
生物制品活性评估是质量控制的关键环节。目前，体外检测是公司和监管机构评估生物活性所采用的主要方法。报告基因检测（RGA）是一种通过易于检测的报告基因来研究基因表达调控和细胞信号转导通路激活的技术。RGA高度依赖药物作用机制，具有高准确性和精密度，且日益受到认可。基于RGA的替代分析方法的应用已成为一种普遍趋势，越来越多的抗体药物采用相应的基于RGA的质量控制方法。当采用RGA技术时，建立稳定表达细胞系对于确保检测在不同条件下的稳定性、可靠性和一致性至关重要。CRISPR/Cas9基因编辑技术介导的位点特异性基因整合能够将外源基因快速、精确地插入特定基因组位点，并实现稳定RGA细胞系的高效构建，这将显著推动生物活性评估方法的进步。
