Host cell proteins in biologics development: Identification, quantitation and risk assessment

  title={Host cell proteins in biologics development: Identification, quantitation and risk assessment},
  author={Xing Wang and Alan K. Hunter and Ned M. Mozier},
  journal={Biotechnology and Bioengineering},
Host cell proteins (HCPs) are those produced or encoded by the organisms and unrelated to the intended recombinant product. Some are necessary for growth, survival, and normal cellular processing whereas others may be non‐essential, simply carried along as baggage. Like the recombinant product, HCPs may also be modified by the host with a number of post‐translational modifications. Regardless of the utility, or lack thereof, HCPs are undesirable in the final drug substance. Though commonly… 

Host cell proteins in biotechnology‐derived products: A risk assessment framework

Factors to be considered in an assessment of risk of residual host cell protein(s) detected and identified in the drug product will lead to an overall risk assessment that informs decision‐making around how to control the levels of host cell proteins.

The future of host cell protein (HCP) identification during process development and manufacturing linked to a risk‐based management for their control

A nascent approach to risk assessment of HCPs based upon such data is described, drawing attention to timeliness in relation to biosimilar initiatives.

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Development of synthetic peptide ligands capable of capturing a broad spectrum of Chinese hamster ovary (CHO) HCPs with a combination of peptide species that allow for advanced mixed-mode binding is described.

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The same preparation of polyclonal anti-HCP antibodies was used to both capture and report antibody for HCP quantitation in the HCP ELISA, so it was hypothesized that even a small fraction of cross-reactivity could cause a measurable HCP signal.

Host cell protein analysis in therapeutic protein bioprocessing – methods and applications

Different methods for the comparison of bioprocessing strategies during scale‐up and purification development are compared and include immunospecific methods, such as ELISA, western blot, and threshold, and non‐specific methods,such as 2D‐DIGE and2D‐HPLC combined with MS.

Host cell protein dynamics in recombinant CHO cells

Understanding the HCP dynamics will in the future help provide a platform to rationally manipulate and engineer and/or select suitable recombinant CHO cell lines and downstream processing steps to limit problematic HCPs.

Profiling the effects of process changes on residual host cell proteins in biotherapeutics by mass spectrometry

An advanced liquid chromatography/mass spectrometry platform was used to identify and quantify residual Escherichia coli host cell proteins (HCPs) in the drug substance (DS) of several peptibodies, demonstrating that HCPs that carry through purification processes to be detectable in DS are not always among those that are the most abundant upstream.

Exploring sample preparation and data evaluation strategies for enhanced identification of host cell proteins in drug products of therapeutic antibodies and Fc-fusion proteins

An improved strategy for evaluating HPLC-MS/MS data of HCP-derived peptides, involving probabilistic protein inference and peptide detection in the absence of fragment ion spectra is described.

Comparison of platform host cell protein ELISA to process‐specific host cell protein ELISA

The evaluation of the suitability of the existing HCP ELISA for monitoring the HCP population in the updated process concluded that the platform H CP ELISA method is superior to the process‐specific HCPELISA method.



Demonstration of Robust Host Cell Protein Clearance in Biopharmaceutical Downstream Processes

The demonstration of robust HCP clearance through this comprehensive strategy can potentially be used to eliminate the need for routine analytical testing or for establishing acceptance criteria for these impurities as well as to demonstrate robust operation of the entire downstream purification process.

Proteomic studies support the use of multi‐product immunoassays to monitor host cell protein impurities

A comparative proteomics study of three independently generated CHO cell lines was performed and results suggest that multi‐product immunoassays are suitable for monitoring host cell proteins in biopharmaceuticals produced in different CHO cell Lines.

Identification of cellular changes associated with increased production of human growth hormone in a recombinant Chinese hamster ovary cell line

A proteomics approach was used to identify the proteins potentially implicated in the cellular response concomitant with elevated production levels of human growth hormone in a recombinant Chinese

Comparative proteomic analysis of GS-NS0 murine myeloma cell lines with varying recombinant monoclonal antibody production rate.

Data reveal both the adaptive responses and molecular mechanisms enabling mammalian cells in culture to achieve high-level recombinant monoclonal antibody production.

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A significant mechanistic finding presented in this work is that HCP contaminants that persist following Protein A capture predominantly comprise species that associate with the product in preference to direct interaction with the chromatographic resin, suggesting that the development of improved column wash techniques to maximize HCP clearance ought to focus on disrupting protein–HCP interactions rather than Protein A–H CP interactions.

Downstream processing of monoclonal antibodies--application of platform approaches.