Next Article in Journal
Targeted Nanobody-Based Molecular Tracers for Nuclear Imaging and Image-Guided Surgery
Previous Article in Journal
A Strategy to Optimize the Generation of Stable Chromobody Cell Lines for Visualization and Quantification of Endogenous Proteins in Living Cells
Previous Article in Special Issue
A RAGE-Targeted Antibody-Drug Conjugate: Surface Plasmon Resonance as a Platform for Accelerating Effective ADC Design and Development
Article Menu

Export Article

Open AccessArticle

Multiplex LC-MS/MS Assays for Clinical Bioanalysis of MEDI4276, an Antibody-Drug Conjugate of Tubulysin Analogue Attached via Cleavable Linker to a Biparatopic Humanized Antibody against HER-2

1
PPD Laboratories, 2244 Dabney Road, Richmond, VA 23230, USA
2
Clinical Immunology and Bioanalysis, Clinical Pharmacology and DMPK, MedImmune LLC, 121 Oyster Point Boulevard, South San Francisco, CA 94080, USA
3
Present address: Pharmacology & Toxicology, CytomX Therapeutics, Inc., 151 Oyster Point Boulevard, Suite 400, South San Francisco, CA 94080, USA
*
Authors to whom correspondence should be addressed.
Antibodies 2019, 8(1), 11; https://doi.org/10.3390/antib8010011
Received: 19 October 2018 / Revised: 6 December 2018 / Accepted: 6 December 2018 / Published: 11 January 2019
(This article belongs to the Special Issue Antibody-Drug Conjugate)
  |  
PDF [1392 KB, uploaded 14 January 2019]
  |  

Abstract

Bioanalysis of complex biotherapeutics, such as antibody-drug conjugates (ADCs), is challenging and requires multiple assays to describe their pharmacokinetic (PK) profiles. To enable exposure-safety and exposure-efficacy analyses, as well as to understand the metabolism of ADC therapeutics, three bioanalytical methods are typically employed: Total Antibody, Antibody Conjugated Toxin or Total ADC and Unconjugated Toxin. MEDI4276 is an ADC comprised of biparatopic humanized antibody attached via a protease-cleavable peptide-based maleimidocaproyl linker to a tubulysin toxin (AZ13599185) with an approximate average drug-antibody ratio of 4. The conjugated payload of MEDI4276 can undergo ester hydrolysis to produce the conjugated payload AZ13687308, leading to the formation of MEDI1498 (de-acetylated MEDI4276). In this report, we describe the development, validation and application of three novel multiplex bioanalytical methods. The first ligand-binding liquid chromatography coupled with tandem mass spectrometry (LBA-LC-MS/MS) method was developed and validated for simultaneous measurement of total antibody and total ADC (antibody-conjugated AZ13599185) from MEDI4276. The second LBA-LC-MS/MS assay quantified total ADC (antibody-conjugated AZ13687308) from MEDI1498. The third multiplex LC-MS/MS assay was used for simultaneous quantification of unconjugated AZ13599185 and AZ13687308. Additional stability experiments confirmed that quantification of the released warhead in the presence of high concentrations of MEDI4276 was acceptable. Subsequently, the assays were employed in support of a first-in-human clinical trial (NCT02576548). View Full-Text
Keywords: Antibody-drug conjugates (ADCs); Hybrid LBA-LC-MS/MS; Multiplexing; Tubulysin; Total ADC; Total Antibody; unconjugated payload; MEDI4276; AZ13599185; AZ13687308 Antibody-drug conjugates (ADCs); Hybrid LBA-LC-MS/MS; Multiplexing; Tubulysin; Total ADC; Total Antibody; unconjugated payload; MEDI4276; AZ13599185; AZ13687308
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Faria, M.; Peay, M.; Lam, B.; Ma, E.; Yuan, M.; Waldron, M.; Mylott, W.R., Jr.; Liang, M.; Rosenbaum, A.I. Multiplex LC-MS/MS Assays for Clinical Bioanalysis of MEDI4276, an Antibody-Drug Conjugate of Tubulysin Analogue Attached via Cleavable Linker to a Biparatopic Humanized Antibody against HER-2. Antibodies 2019, 8, 11.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Antibodies EISSN 2073-4468 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top