Universal Affinity Capture Liquid Chromatography-Mass Spectrometry Assay for Evaluation of Biotransformation of Site-Specific Antibody Drug Conjugates in Preclinical Studies

2019 ◽  
Vol 92 (2) ◽  
pp. 2065-2073 ◽  
Author(s):  
Srikanth Kotapati ◽  
David Passmore ◽  
Sayumi Yamazoe ◽  
Rajesh Kishore Kumar Sanku ◽  
Qiang Cong ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Specific Antibody ◽  
Preclinical Studies ◽  
Liquid Chromatography Mass Spectrometry ◽  
Affinity Capture ◽  
Site Specific ◽  
Drug Conjugates ◽  
Antibody Drug ◽  
Mass Spectrometry Assay

Custom-Designed Affinity Capture LC-MS F(ab′)2 Assay for Biotransformation Assessment of Site-Specific Antibody Drug Conjugates

2016 ◽  
Vol 88 (23) ◽  
pp. 11340-11346 ◽  
Author(s):  
Dian Su ◽  
Carl Ng ◽  
Mehraban Khosraviani ◽  
Shang-Fan Yu ◽  
Ely Cosino ◽  
...  
Keyword(s):  
Specific Antibody ◽  
Affinity Capture ◽  
Site Specific ◽  
Antibody Drug Conjugates ◽  
Drug Conjugates ◽  
Antibody Drug

scholarly journals Insights from native mass spectrometry approaches for top- and middle- level characterization of site-specific antibody-drug conjugates

mAbs ◽  
2017 ◽  
Vol 9 (5) ◽  
pp. 801-811 ◽  
Author(s):  
Thomas Botzanowski ◽  
Stéphane Erb ◽  
Oscar Hernandez-Alba ◽  
Anthony Ehkirch ◽  
Olivier Colas ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Specific Antibody ◽  
Native Mass Spectrometry ◽  
Middle Level ◽  
Site Specific ◽  
Antibody Drug Conjugates ◽  
Drug Conjugates ◽  
Antibody Drug

scholarly journals State-of-the-Art Native Mass Spectrometry and Ion Mobility Methods to Monitor Homogeneous Site-Specific Antibody-Drug Conjugates Synthesis

2021 ◽  
Vol 14 (6) ◽  
pp. 498
Author(s):  
Evolène Deslignière ◽  
Anthony Ehkirch ◽  
Bastiaan L. Duivelshof ◽  
Hanna Toftevall ◽  
Jonathan Sjögren ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Phase ◽  
Ion Mobility ◽  
State Of The Art ◽  
Native Mass Spectrometry ◽  
Site Specific ◽  
Drug Conjugates ◽  
Conjugation Process ◽  
Antibody Drug

Antibody-drug conjugates (ADCs) are biotherapeutics consisting of a tumor-targeting monoclonal antibody (mAb) linked covalently to a cytotoxic drug. Early generation ADCs were predominantly obtained through non-selective conjugation methods based on lysine and cysteine residues, resulting in heterogeneous populations with varying drug-to-antibody ratios (DAR). Site-specific conjugation is one of the current challenges in ADC development, allowing for controlled conjugation and production of homogeneous ADCs. We report here the characterization of a site-specific DAR2 ADC generated with the GlyCLICK three-step process, which involves glycan-based enzymatic remodeling and click chemistry, using state-of-the-art native mass spectrometry (nMS) methods. The conjugation process was monitored with size exclusion chromatography coupled to nMS (SEC-nMS), which offered a straightforward identification and quantification of all reaction products, providing a direct snapshot of the ADC homogeneity. Benefits of SEC-nMS were further demonstrated for forced degradation studies, for which fragments generated upon thermal stress were clearly identified, with no deconjugation of the drug linker observed for the T-GlyGLICK-DM1 ADC. Lastly, innovative ion mobility-based collision-induced unfolding (CIU) approaches were used to assess the gas-phase behavior of compounds along the conjugation process, highlighting an increased resistance of the mAb against gas-phase unfolding upon drug conjugation. Altogether, these state-of-the-art nMS methods represent innovative approaches to investigate drug loading and distribution of last generation ADCs, their evolution during the bioconjugation process and their impact on gas-phase stabilities. We envision nMS and CIU methods to improve the conformational characterization of next generation-empowered mAb-derived products such as engineered nanobodies, bispecific ADCs or immunocytokines.

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