UHPLC Analysis of Orthoclone OKT3 IgG2 Monoclonal Antibody on BIOshell(TM) A400 Protein C4

UHPLC Analysis of Orthoclone OKT3 IgG2 Monoclonal Antibody on BIOshell™ A400 Protein C4

Conditions

column BIOshell A400 Protein C4; 10 cm x 2.1 mm I.D., 3.4 μm particles (66825-U)
column temp. 75 °C
mobile phase [A] 90:10 (0.1% TFA in water): (0.1% TFA in acetonitrile); [B] 10:90 (0.1% TFA in water): (0.1% TFA in acetonitrile)
gradient 0% B to 50%B in 50 min; 50%B to 100% B in 10 min
flow rate 0.2 mL/min
pressure 1650 psi (114 bar)
sample Orthoclone OKT3, 1 mg/mL, 0.05% TFA in water
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). A RPC method was developed that allows for resolution of the main mAb peak from impurities present in the sample. The inset shows the region around the main analyte, and gives an idea on how complex therapeutic mAb samples can be.
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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