慢病毒载体定量检测方法的研究进展

Advances in quantification of lentiviral vectors

摘要:

免疫疗法正在发展成为一种可用于多种恶性肿瘤的有效且侵入性较小的治疗方法。慢病毒载体 (Lentiviral vectors，LVs) 因其能够稳定地整合相对较大的外源DNA，并且有效地转导分裂细胞和非分裂细胞，已在免疫治疗中显现出巨大潜力。临床应用对慢病毒载体具有较高的质量要求，需要对最终产品进行严格的质量控制以保证其纯度、效力和安全性。其中慢病毒载体的定量检测是产品研发和质控的关键环节之一。文中总结了LVs定量检测的现有方法，包括流式细胞术 (Fluorescence activated cell sorter，FACS)、P24酶联免疫法 (P24 enzyme-linked immunosorbent assay，P24 ELISA)、实时荧光定量PCR方法 (Real-time fluorescence quantitative polymerase chain reaction，RT-qPCR)、纳米粒子跟踪分析 (Nanoparticle tracking analysis，NTA)、可调电阻式脉冲传感 (Tunable resistive pulse sensing，TRPS) 和病毒计数仪 (Virus counter，VC)，并对其优缺点进行比较，对发展新的检测方法和存在的挑战进行了展望。

Abstract:

Immunotherapy is becoming an effective and less invasive strategy that can be applied to the treatment of various malignancies. Lentiviral vectors (LVs) have shown great potential in immunotherapy as they can stably integrate relatively large foreign DNA, and effectively transduce dividing and non-dividing cells. Clinical application needs high quality LVs, and therefore strict quality control of the final products is necessary to ensure their purity, efficacy and safety. The quantitative detection of LVs is among the key parts of product development and quality control. In this paper, the existing methods for quantitative detection of LVs are summarized, including fluorescence activated cell sorter (FACS), P24 enzyme-linked immuno sorbent assay (P24 ELISA), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), nanoparticle tracking analysis (NTA), tunable resistive pulse sensing(TRPS) and virus counter(VC).Their advantages and disadvantages are listed, and future development and challenges are discussed.