title: Dialysis: a characterization method of aggregation tendency.
abstract: All researchers immersed in the world of recombinant protein production are in agreement that often the production and purification process of a protein can become a nightmare due to an unexpected behavior of the protein at different protocol stages. Once the protein is purified, scientists know that they still cannot relax. There is a decisive last step missing: performing a protein dialysis in a suitable buffer for subsequent experimental trials. Here is when we can find proteins that precipitate during dialysis by buffer-related factors (ionic strength, pH, etc.), which are intrinsic to each protein and are difficult to predict. How can we find the buffer in which a protein is more stable and with less tendency to precipitate? In this chapter we go over possible factors affecting the protein precipitation tendency during the dialysis process and describe a general dialysis protocol with tricks to reduce protein aggregation. Furthermore, we propose a fast method to detect the most appropriate buffer for the stability of a particular protein, performing microdialysis on a battery of different buffers to measure afterwards precipitation by a colorimetric method, and thus being able to choose the most suitable buffer for the dialysis of a given protein.
authors.affiliation: Departament de Genètica i de Microbiologia, Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, Cerdanyola del Vallès, 08193, Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Pesarrodona
authors.forename: Ugutz
authors.initials: U
authors.lastname: Unzueta
authors.forename: Esther
authors.initials: E
authors.lastname: Vázquez
mesh_terms: D000818:Animals; D002021:Buffers; D003956:Dialysis / Q000379:methods; D006801:Humans; D066329:Protein Aggregates / Q000502:physiology*; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry*
chemical_list: D002021:Buffers; D066329:Protein Aggregates; D011994:Recombinant Proteins