RESUMO
We propose a method for determining the criticality of residual host cell DNA, which is characterized through two attributes, namely the size and amount of residual DNA in biopharmaceutical product. By applying a mechanistic modeling approach to the problem, we establish the linkage between residual DNA and product safety measured in terms of immunogenicity, oncogenicity, and infectivity. Such a link makes it possible to establish acceptable ranges of residual DNA size and amount. Application of the method is illustrated through two real-life examples related to a vaccine manufactured in Madin Darby Canine Kidney cell line and a monoclonal antibody using Chinese hamster ovary (CHO) cell line as host cells.
Assuntos
Biofarmácia/estatística & dados numéricos , DNA/análise , Contaminação de Medicamentos/estatística & dados numéricos , Modelos Estatísticos , Tecnologia Farmacêutica/estatística & dados numéricos , Animais , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/genética , Biofarmácia/normas , Células CHO , Química Farmacêutica , Qualidade de Produtos para o Consumidor , Cricetulus , Interpretação Estatística de Dados , Cães , Guias como Assunto , Humanos , Vacinas contra Influenza/biossíntese , Vacinas contra Influenza/genética , Vacinas contra Influenza/normas , Células Madin Darby de Rim Canino , Controle de Qualidade , Medição de Risco , Tecnologia Farmacêutica/métodos , Tecnologia Farmacêutica/normasRESUMO
PURPOSE: To evaluate Taylor dispersion analysis (TDA) as a novel method for determination of hydrodynamic radius of therapeutic peptides and proteins in non-stressed and stressed formulations and to compare it with dynamic light scattering (DLS). METHODS: The hydrodynamic radius of oxytocin, bovine serum albumin, various monoclonal antibodies (type IgG) and etanercept at concentrations between 0.05 and 50 mg/ml was determined by TDA and DLS. IgGs and etanercept were stressed (elevated temperatures) and analyzed by TDA, DLS and HP-SEC. RESULTS: TDA and DLS were comparable in sizing non-stressed peptides and proteins in a concentration range of about 0.5 to 50 mg/ml. TDA performed well even at lower concentrations, where DLS tends to provide theoretically high values of the Z-average radius. However, because of differences in the detection physics, DLS was more weighted towards the detection of aggregates in stressed formulations than TDA. Advantageously, TDA was also able to size the small peptide oxytocin, which was not feasible by DLS. CONCLUSION: TDA allows the accurate determination of the hydrodynamic radius of peptides and proteins over a wide concentration range, with little interference from excipients present in the sample. It is marginally less sensitive than DLS in detecting size increase for stressed protein samples.