LC/MS Analysis of Reduced SigmaMAb ADC Mimic on BIOshell(TM) IgG 1000 Ã… C4

LC/MS Analysis of Reduced SigmaMAb ADC Mimic 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] 70:30 water (0.1% DFA): acetonitrile (0.1% DFA); [B] 50:50 water (0.1% DFA): acetonitrile (0.1% DFA)
gradient 0% B to 50% B in 50 min
flow rate 0.2 mL/min
pressure 1370 psi (94 bar)
sample ADC mimic, 500 μg/mL, water (0.1% DFA)
injection 1 μL
detector MSD, ESI-(+), 1000 - 17,000 m/z

Description

Analysis Note SigmaMab Antibody Drug Conjugate (ADC) Mimic, is a non-toxic drug mimic that is utilized as a standard for mass spectrometry and high performance liquid chromatography. The ADC Mimic conjugates SigmaMab (MSQC4), an IgG1 monoclonal antibody, to dansyl fluorophores via an LC-SMCC crosslinker. This dansyl-LC-SMCC attachment serves as the substitution for the toxic payload of a real ADC. This application demonstrates the use of this standard in performing middle up/down analysis of an ADC. Resolution of labeled and unlabeled light chain and heavy chain is achieved on the BIOshell IgG 1000 Ã… C4 column.
Featured Industry Pharmaceutical (small molecule)
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
Other Notes The sample was reduced by exposing the sample to 0.1 M ammonium bicarbonate, 8 M guanidine hydrochloride, and 0.1 M dithiobutylamine at room temperature for one hour.
suitability application for LC-MS

Materials

     
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