Ligand-binding mass spectrometry to study biotransformation of fusion protein drugs and guide immunoassay development: strategic approach and application to peptibodies targeting the thrombopoietin receptor.

The AAPS Journal
Michael HallJean W Lee

Abstract

The knowledge of in vivo biotransformation (e.g., proteolysis) of protein therapeutic candidates reveals structural liabilities that impact stability. This information aids the development and confirmation of ligand-binding assays with the required specificity for bioactive moieties (including intact molecule and metabolites) for appropriate PK profiling. Furthermore, the information can be used for re-engineering of constructs to remove in vivo liabilities in order to design the most stable candidates. We have developed a strategic approach of ligand-binding mass spectrometry (LBMS) to study biotransformation of fusion proteins of peptides fused to human Fc ("peptibodies") using anti-human Fc immunoaffinity capture followed by tiered mass spectrometric interrogation. LBMS offers the combined power of selectivity of ligand capture with the specificity and detailed molecular-level information of mass spectrometry. In this paper, we demonstrate the preclinical application of LBMS to three peptibodies, AMG531 (romiplostim), AMG195(linear), and AMG195(loop), that target the thrombopoietin receptor. The data show that ligand capture offers excellent sample cleanup and concentration of intact peptibodies and metabolites for subsequen...Continue Reading

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Citations

Mar 13, 2012·The AAPS Journal·Narendra ChirmuleBernd Meibohm
Apr 10, 2012·The AAPS Journal·Thomayant Prueksaritanont, Cuyue Tang
Mar 1, 2011·Therapeutic Delivery·Jean W Lee, Marian Kelley
Jun 5, 2014·Expert Opinion on Drug Metabolism & Toxicology·Eric EzanFrançois Fenaille
Nov 29, 2013·Journal of Pharmaceutical Sciences·Benjamin Wu, Yu-Nien Sun
Apr 2, 2013·Advanced Drug Delivery Reviews·Eric Ezan
Jul 24, 2012·Drug Discovery Today·Jingjie MoGuodong Chen
Dec 10, 2015·Bioanalysis·Yan J ZhangDharmesh D Desai
Jul 24, 2012·MAbs·Grant ShimamotoChristophe Quéva
Sep 1, 2016·Journal of Immunology Research·Alison SmithFrank-Peter Theil
Jun 21, 2014·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Michael P Hall
Mar 13, 2019·Scientific Reports·Simone SchadtNicole A Kratochwil
Nov 22, 2019·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Simone SchadtChristoph Funk
Dec 22, 2017·Biopolymers·Marco CavacoVera Neves
Apr 23, 2021·Journal of the American Society for Mass Spectrometry·Iain D G Campuzano, Wendy Sandoval

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