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Quality control of protein standards for molecular mass determinations by small‐angle X‐ray scattering
Author(s) -
Akiyama Shuji
Publication year - 2010
Publication title -
journal of applied crystallography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.429
H-Index - 162
ISSN - 1600-5767
DOI - 10.1107/s002188981000138x
Subject(s) - small angle x ray scattering , molecular mass , calibration , scattering , chemistry , macromolecule , fractionation , standard deviation , analytical chemistry (journal) , calibration curve , bovine serum albumin , characterization (materials science) , mass spectrometry , particle (ecology) , chromatography , crystallography , materials science , nanotechnology , optics , physics , detection limit , mathematics , statistics , organic chemistry , biochemistry , oceanography , geology , enzyme
Small‐angle X‐ray scattering (SAXS) is a powerful technique with which to evaluate the size and shape of biological macromolecules in solution. Forward scattering intensity normalized relative to the particle concentration, I (0)/ c , is useful as a good measure of molecular mass. A general method for deducing the molecular mass from SAXS data is to determine the ratio of I (0)/ c of a target protein to that of a standard protein with known molecular mass. The accuracy of this interprotein calibration is affected considerably by the monodispersity of the prepared standard, as well as by the precision in estimating its concentration. In the present study, chromatographic fractionation followed by hydrodynamic characterization is proposed as an effective procedure by which to prepare a series of monodispersed protein standards. The estimation of molecular mass within an average deviation of 8% is demonstrated using monodispersed bovine serum albumin as a standard. The present results demonstrate the importance of protein standard quality control in order to take full advantage of interprotein calibration.