Desenvolvimento de métodos para criopreservação de Proteínas Terapêuticas - Influência da geometria de congelamento na Albumina de Soro Bovino

Rita Garcia Costa

Key information

Authors:

Supervisors:

Andreia Filipa Silvestre Duarte (Andreia Filipa Silvestre Duarte); Miguel Ângelo Joaquim Rodrigues (Miguel Ângelo Joaquim Rodrigues)

Published in

September 29, 2021

Abstract

The cryopreservation is a growing area both in terms of research, development and applications, having great importance in the area of health and pharmaceuticals, duo to is potential to preserve and prolong the activity of proteins and cells. In this work it was study the cryopreservation of bovine serum albumin protein, under two different methods, a unidirectional freezing and a bidirectional freezing. For each method, two formulations were studied. The first solution of BSA include sodium chloride and sucrose and the second solution of BSA just include sodium chloride, both solutions were dissolve in a Tris buffer with pH 7. To assess the impact of freezing in each method, the spatial distribution of protein concentration (cryoconcentration) and the percentage of aggregates were evaluated by size exclusion chromatography and spectroscopy. The formation of insoluble aggregates was detected in the two methods and in the two formulations. The thermophysical properties of the BSA solution were also found to make a simulation in computational fluid dynamics (CFD), using the FreezeSIM software, made available by SmartFreez. In general, the model showed some similarities with the experimental results, being closer to the unidirectional method. It was concluded the formulation with sucrose was a better stabilizer to the protein, duo to its cryoprotectant effect. The percentage of insoluble aggregates was higher in the bidirectional freezing, the unidirectional freezing provides a better stabilization to the protein.