B-221 Quantitation of DNA in Antibody-based and RNA-based Drugs by Quantitative Polymerase Chain Reaction (qPCR)

Abstract Background Current FDA regulation defined acceptance level of 10 ng per dose residual DNA in drug products poses great challenges for accurate quantification DNA, particularly the used high doses or formulated complex matrixes. The purpose this study is to develop and qualify a qPCR method Chinese Hamster Ovary (CHO) antibody-based drugs quantitation human RNA-based drugs. Methods An 8-point standard curve consisting 0.005, 0.05, 0.5, 5, 50, 500, 1000 5000 pg CHO 5-point 0.3, 3, 30, 300 3000 was generated by serial dilutions. antibody sample (300 µg) spiked with 2.5, 10, 50 DNA. RNA (1.6 3 30 samples were analyzed using Real-Time PCR instrument. Commercial test kits (TaqMan™ Probe TaqMan™ Universal Master Mix), sequence specific forward reverse primers used. Results For product, linearity slope R2 are −3.5 1.00, respectively, which met system suitability criteria. accuracy recovery 2.5 pg, µg antibody, 150 formulation buffer, 70%, 76%, 86%, 93% 95%, respectively. precision assay 6%, 7% 2%, QL product.For −3.3 0.3 1.6 RNA, 1.5 113%, 104%, 102%, 97% 88%, 5% product. Conclusion Two methods developed quantify products. One low levels Another