# An Integrated NMR Approach for Evaluating Linker-Payload Conjugation with Monoclonal Antibodies

## TL;DR

This paper introduces a new NMR method to study the structure and properties of antibody-drug conjugates without damaging them.

## Contribution

A novel non-disruptive NMR approach is introduced for characterizing antibody-drug conjugates in a physiological environment.

## Key findings

- The NMR method provides insights into the higher-ordered structure of monoclonal antibodies and ADCs.
- The approach successfully characterized a linker-payload model system using Trastuzumab and a Remdesivir-derived fragment.

## Abstract

Antibody-drug conjugates
(ADCs) are modern biopharmaceuticals that
combine the therapeutic effects of small-molecule drugs with the outstanding
selectivity of monoclonal antibodies (mAbs). Since their introduction
in the biomedical field, research has focused on elucidating the structure,
stability, and mode of action of ADCs. Nevertheless, standard characterization
methods for ADCs heavily rely on disruptive techniques like mass spectrometry
in a non-physiological environment. Here, we present an NMR approach
combining 1H–13C ALSOFAST-HMQC and T2-edited 1H CPMG experiments, which together provide
information on: i. the fingerprint and higher-ordered structure (HOS)
of mAbs and ADCs and ii. the properties of the bound linker-payload
fragment. In this study, we chose Trastuzumab as a well-known mAb
and a Remdesivir-derived fragment as a linker-payload model system
to validate our approach.