Analysis of aggregates of human immunoglobulin G using size-exclusion chromatography, static and dynamic light scattering.
Large aggregates (Mr: 10(6)-10(7) g/mol) of human immunoglobulins are present in extremely small concentrations in IgG preparations (<0.1%). Traces of large protein aggregates cannot be determined by conventional size-exclusion chromatography (SEC) using UV detection due to limitations in sensitivity. The conventional analysis of IgG by SEC is limited to dimers and oligomers. Using light scattering it is possible to determine significant differences concerning the aggregate composition and the extent of protein aggregation in samples of different process steps. Two different pilot preparations were analyzed by SEC with UV and static light scattering detection and compared to dynamic light scattering in the batch mode. The change of large aggregates could be monitored and data were corroborated by dynamic light scattering.
D002850:Chromatography, Gel / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D008027:Light; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity
D007074:Immunoglobulin G