Determination of the molecular size distribution of immunoglobulin G (IgG) in intravenous IgG-albumin formulations by high-performance liquid chromatography.
Recent reports have expressed concern about the safety of intravenous human immunoglobulin G (IgG) preparations. Evidence seems to indicate that aggregates in these formulations are responsible for several adverse reactions in some patients, including anaphylaxis and dyspnea. Therefore, monitoring the molecular size distribution of IgG in these products is essential for ensuring their safety. This paper describes a sensitive and precise two-stage high-performance liquid chromatographic method for determining the molecular size distribution profile of IgG in an intravenous formulation stabilized with albumin. In the first step, all molecular forms of IgG are separated from all molecular forms of albumin by anion-exchange chromatography. In the second stage, a portion of the collected IgG fraction is re-chromatographed by size-exclusion chromatography and separated into its aggregate, dimer, and monomer components. Although some minor losses of aggregate do occur in the procedure, the overall molecular size distribution is not significantly affected. The method described is both time-efficient and accurate, and represents an improvement over existing methods.
Lee
J K
JK
Rorer Central Research, Horsham, PA 19044.
Deluccia
F J
FJ
Kelly
E L
EL
Davidson
C
C
Borger
F R
FR
D000418:Albumins / Q000008:administration & dosage; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006801:Humans; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000032:analysis*; D007275:Injections, Intravenous; D008970:Molecular Weight
D000418:Albumins; D007074:Immunoglobulin G