Sensitive quantification and fragment size assessment of residual host cell DNA using digital PCR

Residual host cell DNA is a process-related impurity that must be controlled for both amount and fragment size in viral vector and gene therapy products. This article describes a digital PCR workflow that combines sensitive absolute quantification of human residual DNA with median fragment size assessment in a single reaction, and presents performance data alongside an independent evaluation using process samples.

What you will learn

01

How a triplex dPCR assay using short (83 bp) and long (345 bp) amplicons at the same locus combines absolute quantification of HEK293 residual DNA with median fragment size assessment in a single reaction

02

Why an extraction-free workflow maintains picogram-level sensitivity (LOQ 4 pg/reaction) and linearity across four orders of magnitude, including directly in conditioned HEK293 media

03

What an independent evaluation with viral process samples showed about skipping DNA isolation, and how optional E1A, E1B, or SV40 multiplexing adds risk-based oncogene detection

Key interests

Residual host cell DNA Digital PCR Fragment size analysis HEK293 Viral vector manufacturing Process-related impurities Extraction-free workflow Oncogene detection Quality control