よくある質問

凍結融解、蒸発、チューブ壁への付着などの影響を避けるため、1回の委託速達サンプル量は50μL以上とすることを推奨します。ゲノム力価、プラスミドDNA残留物、rcAAVなどの検査では10μL以上のサンプルを送付することを推奨します。感染力価検査では20μL以上のサンプルを送付することを推奨します。空殻率検査では5E+13vgのサンプルを送付することを推奨します。

The %RSD of general biochemical methods is usually 15%-25%, and the accepted standard of ddPCR is 10%. In most cases the %RSD for PackGene’s QC ddPCR verification data is lower than 5%, and the %RSD of TCID50 is lower than 10%.

The harvest liquid generated through PackGene’s AAV fast service will be broadly tested for exogenous viral elements as necessary following an evaluation of project characteristics. Both the original harvest liquid and final deliverables for the AAV Fast service will be devoid of such exogenous viral elements.

Yes.

The expected empty shell rate for our AAV Fast Services varies across AAV serotypes. As one example, the expected empty shell rate for AAV9 generated through our AAV Fast Service is lower than 10%.

ウイルスの空殻率は、陰イオンクロマトグラフィーHPLC、分析用超遠心分離法（AUC）、透過型電子顕微鏡（TEM）、CyroTEM、VG力価／カプシド力価など、様々な手法で測定できます。AUC、TEM、CyroTEMは通常、定量的な品質管理測定には適していないため、PackGeneでは陰イオンクロマトグラフィーHPLCを空殻率測定の標準法としています。PackGeneは、陰イオンクロマトグラフィーHPLCで得られた結果の二次検証として、CyroTEMおよびAUC分析サービスも提供しています。

Genome copies per ml (GC/ml) can be determined using primers directed at common vector elements during the early development stage, and thus we use common features such polyA segments or ITRs to determine GC/ml during this stage. However, it is recommended that specific primers targeted at AAV vector transgenes are used for testing the GC of GMP products. Our final fast service titer test is designed to use transgenes specific primers.

The terms genome copies per ml (GC/ml) and viral genomes per ml (vg/ml) are interchangeable and equal in most cases. At PackGene we may test GC by both qPCR and ddPCR. Testing by qPCR involves the use of a calibration standard while ddPCR may use optional reference products. Measurements by qPCR are more likely to be influenced by inter-lab and inter-operator variables, and ddPCR generally shows lower %RSD precision. Typically, GC is determined by qPCR during process exploration phase and for intermediate products while the GC of final products is more often determined by ddPCR.