Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10533274 | Analytical Biochemistry | 2005 | 8 Pages |
Abstract
A method for rapid detection and characterization of reversible associations of macromolecules in solution is presented. A programmable dual-syringe infusion pump is used to introduce a solution of time-varying composition into parallel flow cells for concurrent measurement of laser light scattering at multiple angles and ultraviolet-visible absorbance. An experiment lasting less than 15Â min produces a large and information-rich set of data, consisting of several thousand values of the Rayleigh ratio as a function of solute concentration(s) and scattering angle. Using a novel treatment of the data, the entire data set may be equally rapidly analyzed in the context of models for self-association. Validation experiments conducted on previously characterized nonassociating and self-associating proteins yielded robust values for molecular weights in the range 10-330Â kDa and equilibrium association constants for dimer formation in the range 2Â ÃÂ 103-6Â ÃÂ 105Â Mâ1.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Arun K. Attri, Allen P. Minton,