Detection of Host Cell microprotein impurities In antibody Drug Products

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Oct 4

PMID 39366928

Authors

Ioanna Tzani

Marina Castro-Rivadeneyra

Paul Kelly

Lisa Strasser

Lin Zhang

Martin Clynes

Barry L Karger

Niall Barron

Jonathan Bones

Colin Clarke

Affiliations

Soon will be listed here.

Abstract

Chinese hamster ovary (CHO) cells are used to produce almost 90% of therapeutic monoclonal antibodies (mAbs) and antibody fusion proteins (Fc-fusion). The annotation of non-canonical translation events in these cellular factories remains incomplete, limiting our ability to study CHO cell biology and detect host cell protein (HCP) impurities in the final antibody drug product. We utilised ribosome footprint profiling (Ribo-seq) to identify novel open reading frames (ORFs) including N-terminal extensions and thousands of short ORFs (sORFs) predicted to encode microproteins. Mass spectrometry-based HCP analysis of eight commercial antibody drug products (7 mAbs and 1 Fc-fusion protein) using the extended protein sequence database revealed the presence of microprotein impurities. We present evidence that microprotein abundance varies with growth phase and can be affected by the cell culture environment. In addition, our work provides a vital resource to facilitate future studies of non-canonical translation and the regulation of protein synthesis in CHO cell lines.

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