UHPLC Analysis of Intact IgG4 on BIOshell IgG 1000 Å C4

UHPLC Analysis of Intact IgG4 on BIOshell IgG 1000 Å C4

Conditions

instrument Thermo Ultimate 3000 BioRS
column FPP 300 Å C4, 10 cm x 2.1 mm I.D., 1.7 µm
column BIOshell IgG 1000 Å C4, 10 cm x 2.1 mm I.D., 2.7 µm
mobile phase [A] water (0.1% difluoroacetic acid); [B] acetonitrile (0.1% difluoroacetic acid)
gradient Hold at 22% B for 2 min; 22% B to 52% B in 15 min
flow rate 0.3 mL/min
column temp. 75 °C
detector UV, 215 nm
injection 5.0 µL
sample IgG4, 100 µg/mL, water

Description

Analysis Note Immunoglobulin G4 (IgG4) is the least abundant form of IgG that exists in the human immune system.  However, several diseases are linked to abnormal expression of the protein or due to a low concentration of IgG4.  In addition, several biotherapeutics are based off of this type of IgG.  This application demonstrates the use of the BIOshell IgG 1000 Å C4 column to analyze an intact version of IgG4.  One sharp peak is observed with this column with minimal band broadening relative to a conventional, fully porous particle packed, C4 column. 
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
LiChrosolv is a trademark of Merck KGaA, Darmstadt, Germany
Application No. MUR050618

Materials

     
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