Development of Peptide Ligands for Targeted Capture of Host Cell Proteins from Cell Culture Production Harvests

R Ashton Lavoie; Taufika Islam Williams; R Kevin Blackburn; Ruben G Carbonell; Stefano Menegatti

doi:10.1007/978-1-0716-1186-9_31

Development of Peptide Ligands for Targeted Capture of Host Cell Proteins from Cell Culture Production Harvests

Methods Mol Biol. 2021:2261:489-506. doi: 10.1007/978-1-0716-1186-9_31.

Authors

R Ashton Lavoie¹, Taufika Islam Williams², R Kevin Blackburn³, Ruben G Carbonell^{4

5}, Stefano Menegatti^{6

7}

Affiliations

¹ Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA.
² Molecular Education, Technology, and Research Innovation Center (METRIC), North Carolina State University, Raleigh, NC, USA.
³ Waters Corporation, Morrisville, NC, USA.
⁴ Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA. ruben@ncsu.edu.
⁵ Biomanufacturing Training and Education Center (BTEC), North Carolina State University, Raleigh, NC, USA. ruben@ncsu.edu.
⁶ Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA. smenega@ncsu.edu.
⁷ Biomanufacturing Training and Education Center (BTEC), North Carolina State University, Raleigh, NC, USA. smenega@ncsu.edu.

PMID: 33421010
DOI: 10.1007/978-1-0716-1186-9_31

Abstract

Capture of host cell proteins (HCPs) from cell culture production harvests is critical to ensure the maximum levels specified by international regulatory bodies of product purity for therapeutic monoclonal antibodies (mAbs). Peptide ligands that selectively target the whole spectrum of the HCPs, while letting the mAb product flow through unbound, are an ideal complement to the affinity-based capture step via Protein A chromatography. In this work, we describe the development of HCP-binding peptide ligands, especially focusing on the steps of (1) peptide selection via library screening and (2) quantification of HCP removal via proteomics by mass spectrometry.

Keywords: Flow-through chromatography; Host cell proteins; Mass spectrometry; Monoclonal antibodies; Peptide ligands; Proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CHO Cells
Cricetulus
Enzyme-Linked Immunosorbent Assay
Humans
Ligands
Microscopy, Fluorescence
Peptide Library
Peptides / metabolism*
Protein Binding
Proteins / isolation & purification*
Proteins / metabolism
Proteomics*
Solid Phase Extraction
Tandem Mass Spectrometry

Substances

Ligands
Peptide Library
Peptides
Proteins