Polymerized and polyethylene glycol-conjugated hemoglobins: a globin-based calibration curve for dynamic light scattering analysis.
Dynamic light scattering (DLS) is a technique capable of determining the hydrodynamic radius of proteins. From this parameter, a molecular weight can be assessed provided that an appropriate calibration curve is available. To this goal, a globin-based calibration curve was used to determine the polymerization state of a recombinant hemoglobin-based oxygen carrier and to assess the equivalent molecular weight of hemoglobins conjugated with polyethylene glycol molecules. The good agreement between DLS values and those obtained from gel filtration chromatography is a consequence of the high similarity in structure, shape, and density within the globin superfamily. Moreover, globins and heme proteins in general share similar spectroscopic properties, thereby reducing possible systematic errors associated with the absorption of the probe radiation by the chromophore.
Faggiano
Serena
S
Department of Biochemistry and Molecular Biology, University of Parma, 43124 Parma, Italy.
Ronda
Luca
L
Bruno
Stefano
S
Jankevics
Hanna
H
Mozzarelli
Andrea
A
D000818:Animals; D001802:Blood Substitutes / Q000737:chemistry*; D002138:Calibration; D005914:Globins / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry*; D006736:Horses; D006801:Humans; D008027:Light; D011092:Polyethylene Glycols / Q000737:chemistry*; D012542:Scattering, Radiation
D001802:Blood Substitutes; D006454:Hemoglobins; C109960:PEG-hemoglobin; D011092:Polyethylene Glycols; D005914:Globins