UHPLC Analysis of Orthoclone OKT3 Monoclonal Antibody on BIOshell(TM) IgG 1000 Ã… C4

UHPLC Analysis of Orthoclone OKT3 Monoclonal Antibody on BIOshell™ IgG 1000 Ã… C4

Conditions

column BIOshell IgG 1000 Ã… C4; 10 cm x 2.1 mm I.D., 2.7 μm particles (63288-U)
column temp. 75 °C
mobile phase [A] 90:10 water (0.1% TFA): acetonitrile (0.1% TFA); [B] 10:90 water (0.1% TFA): acetonitrile (0.1% TFA)
gradient 0% B to 50% B in 50 min; 50% B to 100% B in 10 min
flow rate 0.2 mL/min
pressure 1300 psi (89.7 bar)
sample Orthoclone OKT3, 1 g/L, water (0.05% TFA)
injection 1.0 μL
detector UV, 215 nm

Description

Analysis Note Orthoclone OKT3 (common name: muromonab-CD3) is a recombinant monoclonal antibody (mAb) to human CD3. This mAb is used as an immunosuppressant drug given to patients to reduce acute rejection of foreign organs during an organ transplant. This mAb also has the distinction of being the first mAb to be approved for clinical use in humans by the FDA. Due to its use as a therapeutic, the finished drug product needs to be carefully characterized in order to make sure that it is impurity and/or degradant free. One way to characterize these therapeutic mAbs is by reversed-phase chromatography (RPC). Using the BIOshell IgG 1000 Ã… C4 column, resolution of impurites and other modified species was achieved. In addition, the advantages of a larger pore size and utilizing superficially porous particles (SPPs) can be seen in the much improved peak shape of the analyte on the BIoshell IgG column over a 300 Ã…, fully porous particle (FPP) packed competitor column.
Featured Industry Life Science and Biopharma
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
suitability application for UHPLC

Materials

     
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