Application of Quality by Design in a Commercialized Lyophilized Vaccine

The pharmaceutical industry has being implementing regulatory practices to assure that consumers obtain products with quality, safety, and efficacy. The use of Quality by Design (QbD) for products on development has increased though the years to avoid issues related to quality parameters and has been suggested by Regulatory Agencies to standardize globally the documentation for the registration of new products. Although the concepts of QbD gain importance, it is still not a widespread practice to existing systems and products already on the market. This work aims to propose a case study with a vaccine using QbD concepts on the lyophilization unit operation production step to provide robustness and increase efficiency, leading to a lyophilization cycle time reduction. To this end, a reverse way of the use of QbD principles were applied based on historical batches database, down scale experiments, and finally in industrial scale to establish new boundaries in the lyophilization cycle. Experimental batches samples were analyzed through accelerated and real time stability study. At the end, this case became a possibility to establish new ranges to lyophilization process predicting risks and assure robustness to this production step with the maintenance of quality and safety of vaccine

Análise de Risco: estado da arte da metodologia Hazop generalizada, aplicações e perspectivas na indústria de processos

Introduction: The Hazard and Operability Study is considered a feasible tool to assess risks, where complex technologies, require new strategies to guarantee efficiency, safety, and quality of products. Objective: To perform a Hazop publications review, to establish the state of the art, current procedures and perspectives in the pharmaceutical industry. Method: Hazop methodology and improvements to satisfy actual needs were structured. Subsequently, its application and integration with other risk tools, and experts systems, were analyzed to define the current approach and future perspectives. Results: The review allowed the understanding where models, simulations and specialized software offered adequate support to assess risk in current complex processes. In addition, an efficient definition of causes and consequences depends of expert systems, where simulations acquire experience through the creation of databases, reducing the need of specific process knowledge, which is a typical limitation of the conventional Hazop methodology. Conclusions: A review of the Hazop stateof- the-art highlighted the importance to assess risks within the process industry. However, the use of new technologies designed to meet regulatory affairs to guarantee safety and quality principles would require the ongoing improvement of the Hazop methodology, restricting the dependence of specialists, and increasing the use of expert systems.Introducción: El Estudio de Riesgos y Operabilidad se considera una herramienta factible para evaluar los riesgos, cuando las tecnologías complejas requieren nuevas estrategias para garantizar la eficiencia, la seguridad y la calidad de los productos. Objetivo: realizar una revisión de las publicaciones de Hazop, para establecer el estado del arte, los procedimientos actuales y las perspectivas en la industria farmacéutica. Método: fue estructurada la metodología Hazop y las mejoras para satisfacer las necesidades reales. Posteriormente, se analizó su aplicación e integración con otras herramientas de riesgo y sistemas de expertos para definir el enfoque actual y las perspectivas futuras. Resultados: la revisión permitió comprender dónde los modelos, las simulaciones y el software especializado ofrecían el soporte adecuado para evaluar el riesgo en los procesos complejos actuales. Además, una definición eficiente de causas y consecuencias depende de los sistemas expertos, donde las simulaciones adquieren experiencia a través de la creación de bases de datos, lo que reduce la necesidad de un conocimiento específico del proceso, que es una limitación típica de la metodología convencional Hazop. Conclusiones: una revisión del estado del arte de Hazop resaltó la importancia de evaluar los riesgos dentro de la industria de procesos. Sin embargo, el uso de nuevas tecnologías diseñadas para cumplir con los asuntos regulatorios para garantizar los principios de seguridad y calidad requeriría la mejora continua de la metodología Hazop, restringiendo la dependencia de especialistas y aumentando el uso de sistemas expertos.Título PT: Análise de Risco: estado da arte da metodologia Hazop generalizada, aplicações e perspectivas na indústria de processos Introdução: O Estudo de Perigos e Operabilidade (Hazop) é considerado uma ferramenta para avaliação de riscos, na qual tecnologias complexas exigem novas estratégias para garantir a eficiência, a segurança e a qualidade dos produtos. Objetivo: Realizar uma revisão de publicações do Hazop, para estabelecer o estado da arte, os procedimentos e as suas perspectivas na indústria farmacêutica. Método: O procedimento Hazop e suas adequações para satisfazer as necessidades atuais foram estruturados. Posteriormente, aplicações e integração com outras ferramentas de risco e sistemas expertos foram analisadas para definir a abordagem atual e perspectivas futuras. Resultados: A revisão permitiu a compreensão de que modelos, simulações e software especializado oferecem suporte para avaliar riscos em processos complexos. Adicionalmente, a correta definição de causas e consequências depende do uso de sistemas expertos, cujas simulações adquirem experiência através da criação de bancos de dados, reduzindo a necessidade de conhecimento específico do processo, que é uma limitação da metodologia Hazop convencional. Conclusões: A revisão do estado da arte do Hazop destacou a importância de avaliar riscos dentro da indústria de processos. No entanto, novas tecnologias utilizadas para atender quesitos regulatórios de segurança e qualidade precisam da melhoria contínua da metodologia Hazop, reduzindo a dependência de especialista por meio do uso de sistemas especializados

Kinetoplastid membrane protein-11 is present in promastigotes and amastigotes of Leishmania amazonensis and its surface expression increases during metacyclogenesis

Kinetoplastid membrane protein-11 (KMP-11), a protein present in all kinetoplastid protozoa, is considered a potential candidate for a leishmaniasis vaccine. A suitable leishmaniasis vaccine candidate molecule must be expressed in amastigotes, the infective stage for mammals. However, the expression of KMP-11 in Leishmania amastigotes has been a subject of controversy. We evaluated the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, of Leishmania amazonensis by immunoblotting, flow cytometry and immunocytochemistry, using a monoclonal antibody against KMP-11. We found that KMP-11 is present in promastigotes and amastigotes. In both stages, the protein was found in association with membrane structures (at the cell surface, flagellar pocket and intracellular vesicles). More importantly, its surface expression is higher in amastigotes than in promastigotes and increases during metacyclogenesis. The increased expression of KMP-11 in metacyclic promastigotes, and especially in amastigotes, indicates a role for this molecule in the parasite relationship with the mammalian host. The presence of this molecule in amastigotes is consistent with the previously demonstrated immunoprotective capacity of vaccine prototypes based on the KMP-11-coding gene and the presence of humoral and cellular immune responses to KMP-11 in Leishmania-infected humans and animals