A generic approach for simultaneous measurements of total antibody and cleavable antibody-conjugated drug by LC/MS/MS

Ling Xu; Laura E Packer; Chao Li; Kojo Abdul-Hadi; Petter Veiby

doi:10.1016/j.ab.2017.08.024

A generic approach for simultaneous measurements of total antibody and cleavable antibody-conjugated drug by LC/MS/MS

Anal Biochem. 2017 Nov 15:537:33-36. doi: 10.1016/j.ab.2017.08.024. Epub 2017 Sep 1.

Authors

Ling Xu¹, Laura E Packer², Chao Li³, Kojo Abdul-Hadi⁴, Petter Veiby⁵

Affiliations

¹ Mersana Therapeutics Inc., 840 Memorial Drive, Cambridge, MA 02139, USA; Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address: [email protected].
² Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address: [email protected].
³ Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address: [email protected].
⁴ Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address: [email protected].
⁵ Millennium Pharmaceuticals, Inc., A Wholly Owned Subsidiary of Takeda Pharmaceutical Company Limited, 35 Landsdowne Street, Cambridge, MA 02139, USA. Electronic address: [email protected].

PMID: 28867503
DOI: 10.1016/j.ab.2017.08.024

Abstract

The current industry practice for antibody-drug conjugate (ADC) bioanalysis includes quantification of total antibody and antibody-conjugated drug. Here, we report a novel 2-in-1 approach for measuring total antibody and protease-cleavable conjugated drug Monomethyl Auristatin E (MMAE) concurrently. This allows for the determination of the DAR (Drug Antibody Ratio) for in vivo samples, with a 3-orders linear range based on total antibody concentration from 0.1 to 100 μg/mL. Our generic, concurrent method has been cross-validated with the previously established methods in an animal study. This novel approach is applicable to all human IgG1 ADCs with papain cleavable conjugated drug in preclinical studies.

Keywords: Antibody quantification; Antibody-conjugated drug; Antibody-drug conjugates; Drug Antibody Ratio; MMAE.

MeSH terms

Antibodies / analysis*
Antibodies / chemistry
Chromatography, High Pressure Liquid*
Humans
Immunoconjugates / analysis*
Immunoconjugates / metabolism
Immunoglobulin G / chemistry
Immunoglobulin G / metabolism
Oligopeptides / analysis
Oligopeptides / chemistry
Papain / metabolism
Tandem Mass Spectrometry*

Substances

Antibodies
Immunoconjugates
Immunoglobulin G
Oligopeptides
Papain
monomethyl auristatin E