Liquid-chromatography-coupled SAXS for accurate sizing of aggregating proteins.
Small-angle X-ray scattering and size-exclusion chromatography have been combined within a unified experimental set-up to obtain molecular size information. Besides providing simultaneous corroborative data bearing on the same question from two distinct experimental techniques, passing the samples over a gel filtration column immediately prior to illumination by X-rays provides both a more homogeneous sample and a continuous set of data as the concentration is extrapolated to zero. This greatly facilitates analysis of data from oligomerizing or aggregating proteins and increases the reliability of the results.
Mathew
Elizabeth
E
Illinois Institute of Technology, 3101 South Dearborn Street, Chicago, IL 60616, USA.
Mirza
Ahmed
A
Menhart
Nick
N
D001665:Binding Sites; D002850:Chromatography, Gel / Q000379:methods*; D045424:Complex Mixtures / Q000032:analysis / Q000737:chemistry; D045304:Cytochromes c / Q000032:analysis / Q000737:chemistry; D019281:Dimerization; D046911:Macromolecular Substances; D010316:Particle Size; D010958:Plasminogen / Q000032:analysis / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014961:X-Ray Diffraction / Q000379:methods*
D045424:Complex Mixtures; D046911:Macromolecular Substances; D011506:Proteins; D010958:Plasminogen; D045304:Cytochromes c