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Open AccessArticle

Quantification of an Antibody-Conjugated Drug in Fat Plasma by an Affinity Capture LC-MS/MS Method for a Novel Prenyl Transferase-Mediated Site-Specific Antibody–Drug Conjugate

1
College of Pharmacy and Institute of Drug Research and Development, Chungnam National University, Daejeon 34134, Korea
2
LegoChemBiosciences, Inc. 8-26 Munpyeongseo-ro Daedeok-gu Daejeon 34302, Korea
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Molecules 2020, 25(7), 1515; https://doi.org/10.3390/molecules25071515 (registering DOI)
Received: 5 March 2020 / Revised: 25 March 2020 / Accepted: 25 March 2020 / Published: 26 March 2020
The novel prenyl transferase-mediated, site-specific, antibody–drug conjugate LCB14-0110 is comprised of a proprietary beta-glucuronide linker and a payload (Monomethyl auristatin F, MMAF, an inhibitor for tubulin polymerization) attached to human epidermal growth factor receptor 2 (HER2)-targeting trastuzumab. A LC-MS/MS method was developed to quantify the antibody-conjugated drug (acDrug) for in vitro linker stability and preclinical pharmacokinetic studies. The method consisted of affinity capture, enzymatic cleavage of acDrug, and LC-MS/MS analysis in the positive ion mode. A quadratic regression (weighted 1/concentration2), with the equation y = ax2 + bx + c, was used to fit calibration curves over the concentration range of 19.17~958.67 ng/mL for acDrug. The qualification run met the acceptance criteria of ±25% accuracy and precision values for quality control (QC) samples. The overall recovery was 42.61%. The dilution integrity was for a series of 5-fold dilutions with accuracy and precision values ranging within ±25%. The stability results indicated that acDrug was stable at all stability test conditions (short-term: 1 day, long-term: 10 months, Freeze/Thaw (F/T): 3 cycles). This qualified method was successfully applied to in vitro linker stability and pharmacokinetic case studies of acDrug in rats. View Full-Text
Keywords: antibody-conjugated drug; LC-MS/MS; bioanalysis; antibody–drug conjugate; beta-glucuronidase antibody-conjugated drug; LC-MS/MS; bioanalysis; antibody–drug conjugate; beta-glucuronidase
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Lee, B.; Park, M.-H.; Byeon, J.-J.; Shin, S.-H.; Choi, J.; Park, Y.; Park, Y.-H.; Chae, J.; Shin, Y.G. Quantification of an Antibody-Conjugated Drug in Fat Plasma by an Affinity Capture LC-MS/MS Method for a Novel Prenyl Transferase-Mediated Site-Specific Antibody–Drug Conjugate. Molecules 2020, 25, 1515.

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