Determination of carbohydrate contents from excess light scattering.
In the light scattering technique, glycosylation gives rise to excess light scattering for glycoproteins. Assuming additivity of refractive index and using an appropriate refractive index increment for carbohydrate, one can determine the degree of glycosylation from the excess light scattering. Here we have used size-exclusion chromatography in combination with online light scattering, UV absorbance, and refractive index. The results show that the technique accurately determines the carbohydrate content of recombinant stem cell factor.
Arakawa
T
T
Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, California 91360, USA. tarakawa2@aol.com
Wen
J
J
D000818:Animals; D002241:Carbohydrates / Q000032:analysis*; D002417:Cattle; D002850:Chromatography, Gel; D006224:Cricetinae; D006023:Glycoproteins / Q000737:chemistry; D010047:Ovalbumin / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012031:Refractometry / Q000379:methods*; D012542:Scattering, Radiation; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D019089:Stem Cell Factor / Q000737:chemistry
D002241:Carbohydrates; D006023:Glycoproteins; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D012710:Serum Albumin, Bovine; D010047:Ovalbumin