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将数字和实时PCR平台进行比较,用于在制造过程中检测剩余的iPSC和产生病毒的细胞

Un Na Koh¹, Si-Keun Lim²

¹Department of Forensic Sciences, Sungkyunkwan University, Suwon 16419, Gyeonggi-do, Republic of Korea.

Analytical biochemistry

|January 21, 2026

概括

数字PCR (dPCR) 提供了比qPCR更一致的量化,用于检测细胞疗法制造中的痕迹残留细胞. 这种数字PCR方法显示变异性降低和变化系数降低,确保可靠的患者安全.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
细胞疗法细胞疗法

背景情况:

数字PCR (dPCR) 对于细胞治疗质量控制 (QC) 中绝对核酸量化至关重要.
检测诸如诱导多能干细胞 (iPSC) 和病毒产生细胞等残留细胞的痕迹对于患者的安全至关重要.
定量PCR (qPCR) 是一种常见的方法,但其在低度的性能需要与dPCR进行比较.

研究的目的:

将两个数字PCR (dPCR) 平台的分析性能与qPCR进行比较,用于检测细胞疗法制造中的残留细胞.
评估qPCR和dPCR在广泛的目标度范围内的灵敏度,线性和一致性.
确定dPCR适用于可靠量化QC应用中的低丰度目标的适用性.

主要方法:

将qPCR (Bio-Rad CFX96 Opus) 与两个dPCR系统 (Bio-Rad QX200,QIAGEN QIAcuity) 进行比较.
使用多能性标记物 (TDGF1,OCT4,NANOG) 分析含有iPSC和分化细胞 (CM,NPC) 的样本.
在CAR-T制剂中量化产生病毒的293T细胞,使用连续稀释 (1:1到1:10^6) 的lentiviral序列 (gag,VSVG).

主要成果:

在大多数稀释中,dPCR平台和qPCR都表现出可比的灵敏度和线性.
与dPCR相比,qPCR在低模板度下显示了更频繁的信号损失.
dPCR显示变异性降低和变化系数 (CV) 较低,这表明在低目标水平下,量化更稳定.

结论:

与qPCR相比,dPCR在稀释范围内提供了更一致的量化,特别是在低目标水平时.
这两种dPCR系统的分析性能相似,支持它们适用于细胞治疗中的QC.
dPCR增强了低丰度残留细胞的可靠检测,有助于提高细胞疗法制造中的患者安全.

关键词:

在CAR-T细胞中.细胞疗法是一种细胞疗法.数字PCR是数字的PCR.滴滴数码PCR可以进行.纳米板数字PCR技术的使用.这就是 iPSC 的意义.

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