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 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.
Toledo-Rubio
Verónica
V
CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Spain.
Vazquez
Esther
E
Platas
Gemma
G
Domingo-Espín
Joan
J
Unzueta
Ugutz
U
Steinkamp
Elke
E
García-Fruitós
Elena
E
Ferrer-Miralles
Neus
N
Villaverde
Antonio
A
D001681:Biological Assay / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism*; D017551:Microdialysis / Q000379:methods*; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D012995:Solubility; D013997:Time Factors
D049452:Green Fluorescent Proteins