Pontos críticos na liofilização dos componentes do selante de fibrina heterólogo
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Data
2021-08-30
Autores
Mennocchi, Juliana de Barros
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Universidade Estadual Paulista (Unesp)
Resumo
RESUMO O Selante Heterólogo de Fibrina (SHF) foi desenvolvido com tecnologia 100% nacional em uma Universidade Pública Brasileira sendo um bioproduto derivado de um hemocomponente rico em fibrinogênio extraído do sangue de búfalos e de uma enzima isolada e purificada do veneno da cascavel Sulamericana. Diferentemente dos selantes de fibrina comercialmente disponíveis, não possui derivados de sangue humano em sua composição, adicionando enormes vantagens a seu uso. Um dos grandes desafios é aumentar o seu tempo de prateleira que é de dois anos e a sua apresentação que é congelado a - 20 oC. Desta maneira, o objetivo deste projeto foi padronizar a técnica de liofilização dos componentes do (SHF) estudando as temperaturas críticas envolvidas no processo. Para isso, diversos parâmetros foram avaliados como os pontos de Congelamento, Transição vítrea, Vitrificação, Ponto Eutético e Temperatura de Colapso. Por meio da avaliação do processo de liofilização utilizando o DSC Q20 com acessório de resfriamento RCS 90 e do FDM – Freezer Drying Microscopy, conseguimos identificar pontos de temperaturas envolvidas no processo de liofilização do crioprecipitado e da giroxina. Os resultados são complementares e, portanto, pudemos mapear o processo de liofilização completa dos componentes do SHF e recomendamos também a necessidade de utilização de tampões e estabilizantes em uma das técnicas já que encontramos duas temperaturas de transição vítrea para a giroxina. Já o crioprecipitado mostrou resultados mais adequados mesmo em sua composição natural. Espera-se com a aplicação dessa técnica, viabilizar o maior escalonamento do produto, podendo, portanto, utilizá-lo amplamente para outras aplicações.
ABSTRACT The Fibrin Heterologous Sealant (SHF) was developed with 100% national technology at a Brazilian Public University and is a bioproduct derived from a blood component rich in buffalo fibrinogen and an enzyme isolated and purified from the venom of the South American rattlesnake. Unlike commercially available fibrin sealants, it does not contain human blood derivatives in its composition, adding enormous advantages to its use. One of the biggest challenges is to increase its shelf life which is two years and its presentation which is frozen at - 20 oC. Thus, the objective of this project was to standardize the freeze-drying technique for the components of (SHF) by studying the critical temperatures involved in the process. For that, several parameters were evaluated, such as the Freezing points, Glass transition, Vitrification, Eutectic point and Collapse Temperature. Through the evaluation of the lyophilization process using the DSC Q20 with cooling accessory RCS 90 and the FDM - Freezer Drying Microscopy, we were able to identify points of temperatures involved in the freeze-drying process of cryoprecipitate and gyroxin. The results are complementary and, therefore, we were able to map the complete lyophilization process of the components of the SHF and we also recommend the need to use buffers and stabilizers in one of the techniques since we found two glass transition temperatures for gyroxin. Cryoprecipitate showed more adequate results even in its natural composition. It is hoped with the application of this technique, to make the product more scalable and, therefore, able to use it widely for other applications.
ABSTRACT The Fibrin Heterologous Sealant (SHF) was developed with 100% national technology at a Brazilian Public University and is a bioproduct derived from a blood component rich in buffalo fibrinogen and an enzyme isolated and purified from the venom of the South American rattlesnake. Unlike commercially available fibrin sealants, it does not contain human blood derivatives in its composition, adding enormous advantages to its use. One of the biggest challenges is to increase its shelf life which is two years and its presentation which is frozen at - 20 oC. Thus, the objective of this project was to standardize the freeze-drying technique for the components of (SHF) by studying the critical temperatures involved in the process. For that, several parameters were evaluated, such as the Freezing points, Glass transition, Vitrification, Eutectic point and Collapse Temperature. Through the evaluation of the lyophilization process using the DSC Q20 with cooling accessory RCS 90 and the FDM - Freezer Drying Microscopy, we were able to identify points of temperatures involved in the freeze-drying process of cryoprecipitate and gyroxin. The results are complementary and, therefore, we were able to map the complete lyophilization process of the components of the SHF and we also recommend the need to use buffers and stabilizers in one of the techniques since we found two glass transition temperatures for gyroxin. Cryoprecipitate showed more adequate results even in its natural composition. It is hoped with the application of this technique, to make the product more scalable and, therefore, able to use it widely for other applications.
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Palavras-chave
Liofilização, Proteínas, Lyophilization, Bioprodutos, Selante de fibrina, Proteins, Bioproducts, Fibrin sealant