A simple LC/MRM-MS-based method to quantify free linker-payload in antibody-drug conjugate preparations.
Antibody-drug conjugates represent a growing class of biologic drugs that use the targeted specificity of an antibody to direct the localization of a small molecule drug, often a cytotoxic payload. After conjugation, antibody-drug conjugate preparations typically retain a residual amount of free (unconjugated) linker-payload. Monitoring this free small molecule drug component is important due to the potential for free payload to mediate unintended (off-target) toxicity. We developed a simple RP-HPLC/MRM-MS-based assay that can be rapidly employed to quantify free linker-payload. The method uses low sample volumes and offers an LLOQ of 10nM with 370pg on column. This analytical approach was used to monitor free linker-payload removal during optimization of the tangential flow filtration manufacturing step.
Catalent Biologics, 5703 Hollis Street, Emeryville, CA 94608, United States.
Wesley
W
Zmolek
Catalent Biologics, 5703 Hollis Street, Emeryville, CA 94608, United States.
Stefanie
S
Bañas
Catalent Biologics, 5703 Hollis Street, Emeryville, CA 94608, United States.
Robyn M
RM
Barfield
Catalent Biologics, 5703 Hollis Street, Emeryville, CA 94608, United States.
David
D
Rabuka
Catalent Biologics, 5703 Hollis Street, Emeryville, CA 94608, United States. Electronic address: penelope.drake@catalent.com.
Penelope M
PM
Drake
D002853:Chromatography, Liquid / Q000379:methods*; D018796:Immunoconjugates / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000737:chemistry*
D018796:Immunoconjugates; D004364:Pharmaceutical Preparations
ADC; Antibody-drug conjugate; Free drug; MRM; Multiple-reaction monitoring