673. Establishment of a National Reference Standard for Reverse Transcriptase Activity Assay and Its Applicability in Quality Control of Gene therapy Products
Year: 2015
Type: article
Abstract: The reverse transcriptase (RT) activity assay is widely used in the quality control of biological products including gene therapy products for it can detect all infectious retroviruses in biological starting materials, cell banks and final products. But the lack of reference standard for RT activity results in significant systemic error and incomparability between laboratories. To standardize RT activity assay, this study established a sensitive real time PCR and developed the first national reference standard, which was prepared from a raw material of recombinant moloney murine leukemia virus reverse transcriptase. Our tests indicated that the candidate reference standard met relevant requirements of the national reference standard. The activity value of candidate RT standard was determined as 26.4 U per ampoule by a collaborative calibration among four laboratories, with the within-laboratory geometric coefficients of variation (GCVs) ranging from 2.7% to 9.6% and the between-laboratory GCV values being 9.8%. The thermal accelerated stability test showed that no loss of activity was observed when the candidate RT standard was stored at 4°C, 25°C and 37 °C compared with -20°C even after 9 months. The applicability study showed that the cell bank and viral bank of an adenovirus gene therapy product were positive and the final products were negative. As a conclusion, the candidate preparation was suitable to serve as a Chinese national standard for RT activity assay and can be used in the quality control of gene therapy products.
Source: Molecular Therapy
Authors: Yonghong Li, Chen Wang, Lei Yu, Chunming Rao, Junzhi Wang
Institution: National Institutes for Food and Drug Control
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W2561904267