Protein aggregation and soluble aggregate formation screened by a fast microdialysis assay

Protein aggregation is a major obstacle in recombinant protein production as it reduces the yield of soluble polypeptides. Also, the formation of aggregates occurring in the soluble fraction is more common than formerly expected, and the prevalence of these entities might significantly affect the av...

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Bibliographic Details
Authors: Toledo-Rubio, Verónica, Platas, Gemma, Domingo-Espin, Joan|||0000-0001-8149-0803, Unzueta Elorza, Ugutz|||0000-0001-5119-2266, Steinkamp, Elke, Garcia-Fruitos, Elena|||0000-0001-7498-4864, Ferrer-Miralles, Neus|||0000-0003-2981-3913, Villaverde, Antonio|||0000-0002-2615-4521
Format: article
Publication Date:2010
Country:España
Institution:Universitat Autònoma de Barcelona
Repository:Dipòsit Digital de Documents de la UAB
Language:English
OAI Identifier:oai:ddd.uab.cat:137805
Online Access:https://ddd.uab.cat/record/137805
https://dx.doi.org/urn:doi:10.1177/1087057110363911
Access Level:Open access
Keyword:Protein aggregation
Nanoparticles
Dynamic light scattering
Proto-aggregates
Soluble aggregates
Description
Summary:Protein aggregation is a major obstacle in recombinant protein production as it reduces the yield of soluble polypeptides. Also, the formation of aggregates occurring in the soluble fraction is more common than formerly expected, and the prevalence of these entities might significantly affect the average quality of the soluble protein species. Usually, the formation of soluble aggregates remains unperceived because analytical methods such as dynamic light scattering are not routinely applied as quality control procedures. The authors have developed a methodologically simple and fast procedure, based on microdialysis and image processing, that reveals the aggregation tendency of a given protein in a specific environment. Because they also show a good correlation between macroscopic aggregation and soluble aggregate formation, the microdialysis approach also permits an estimation of the occurrence of soluble aggregates.