Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses

Catherine E M Hogwood; Lesley M Chiverton; C Mark Smales

doi:10.1007/978-1-4939-6972-2_16

Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses

Methods Mol Biol. 2017:1603:243-250. doi: 10.1007/978-1-4939-6972-2_16.

Authors

Catherine E M Hogwood¹, Lesley M Chiverton¹, C Mark Smales²

Affiliations

¹ Industrial Biotechnology Centre and School of Biosciences, University of Kent, Canterbury, Kent, UK.
² Industrial Biotechnology Centre and School of Biosciences, University of Kent, Canterbury, Kent, UK. C.M.Smales@kent.ac.uk.

PMID: 28493135
DOI: 10.1007/978-1-4939-6972-2_16

Abstract

Host cell protein content during bioprocessing of biotherapeutic proteins generated from cultured Chinese hamster ovary (CHO) cells is typically measured using immunological and gel-based methods. Estimation of HCP concentration is usually undertaken using Enzyme-Linked ImmunoSorbent Assays (ELISA), while estimation of HCP clearance/presence can be achieved by comparing 2D-PAGE images of samples and by undertaking western blotting of 2D-PAGE analyzed samples. Here, we describe the analyses of HCP content using these methodologies.

Keywords: 2D-PAGE; ELISA; Host cell proteins (HCPs); Western blotting.

MeSH terms

Animals
Biotechnology / methods*
Blotting, Western
CHO Cells
Cricetinae
Cricetulus
Electrophoresis, Gel, Two-Dimensional / methods*
Enzyme-Linked Immunosorbent Assay / methods*
Humans
Proteins / analysis*
Proteins / metabolism
Recombinant Proteins / metabolism*

Substances

Proteins
Recombinant Proteins