Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jun 1:1146:122075.
doi: 10.1016/j.jchromb.2020.122075. Epub 2020 Mar 31.

UPLC-based assay to assess the hydrophobicity of Antibody-Drug Conjugate (ADC) payloads

Ilona Pysz  1 , Paul J M Jackson  2 , David J Barlow  3 , Khondaker Miraz Rahman  1 , David E Thurston  4
Affiliations

UPLC-based assay to assess the hydrophobicity of Antibody-Drug Conjugate (ADC) payloads

Ilona Pysz et al. J Chromatogr B Analyt Technol Biomed Life Sci. .

Abstract

Antibody-Drug Conjugates (ADCs) consist of antibodies attached to cytotoxic small molecules or biological agents (i.e., payloads) through chemical linkers which may be cleavable or non-cleavable. The development of new ADCs is challenging, particularly the process of attaching the linker-payload construct to the antibody (i.e., the conjugation process). One of the major problems associated with conjugation is high hydrophobicity of the payload which can lead to low yields of the ADC through aggregation and/or lower than desired Drug-Antibody Ratios (DARs). We report here a UPLC-based assay that can be used to study the physicochemical properties of ADC payloads at an early stage of development, and to provide information on whether the hydrophilic-hydrophobic balance is suitable for conjugation or further physicochemical optimization is required. The assay is relatively simple to establish and should be of use to those working in the ADC area.

Keywords: ADC payload; Aggregation; Analytical method development; Antibody Drug Conjugate (ADC); Conjugation; High-Performance Liquid Chromatography (HPLC); Hydrophobicity; Ultra-Performance Liquid Chromatography (UPLC).

PubMed Disclaimer

Conflict of interest statement

Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: IP has share options and PJMJ, KMR and DET have founding stock in Femtogenix Ltd, the company supplying one of the experimental ADC payloads evaluated in this study. IP and PJMJ are fully employed by Femtogenix Ltd, and KMR and DET have consultancies with the company. PJMJ, KMR and DET are named inventors on patents filed and owned by the company.

MeSH terms

LinkOut - more resources