Host cell contaminant protein assay development for recombinant biopharmaceuticals

Eaton, Leslie C.

doi:10.1016/0021-9673(94)01249-e

Cited by 89 publications

(69 citation statements)

References 37 publications

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“…During the developmental process, removal of CHO cell-derived proteins from the recombinant product is monitored using multiproduct immunoassays. Such immunoassays are developed by raising antibodies to a single CHO protein preparation, usually from nonproducing CHO cell lines (3,4).…”

Section: Introductionmentioning

confidence: 99%

Proteomic Profiling of Secreted Proteins from CHO Cells Using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry

et al. 2008

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Chinese hamster ovary (CHO) cells are the most commonly used host cell line for the production of recombinant biopharmaceuticals. These biopharmaceuticals are typically secreted from CHO cells and purified from harvested cell culture media. The purpose of this study was to investigate changes in the secreted proteome of CHO cells over the various stages of the growth cycle using Surface Enhanced Laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). Conditioned media samples were collected each day over a 6 day growth period from CHO-K1 cells grown in low serum (0.5% FBS) conditions in monolayer culture. Samples were profiled on a number of ProteinChip arrays with different chromatographic surfaces. From this study, 24 proteins were found to be differentially regulated at different phases of the growth cycle in CHO-K1 cells, when profiled on two chromatographic surfaces, Q10 (anionic) and IMAC30 (metal affinity) ProteinChip arrays.

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Section: Introductionmentioning

confidence: 99%

Proteomic Profiling of Secreted Proteins from CHO Cells Using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry

et al. 2008

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“…Many analytical technologies have been used for the detection, identification, quantitation, and risk assessment of HCPs (Briggs and Panfili, 1991;Eaton, 1995;Hoffman, 2000). It is expedient to select the right combination of technologies for any specific bioprocess, and most importantly, to understand the advantage and limitation of each technology so that an optimal strategy can be developed for a bioprocess-specific HCP evaluation.…”

Section: Analytical Technology Overviewmentioning

confidence: 99%

“…Preparing the immunogen carefully is essential since it will also be used as a raw material to standardize the quantitation antibodies. Many different approaches have been employed to raise antibodies to HCP antigens (Briggs and Panfili, 1991;Eaton, 1995;Thalhamer and Freund, 1984). Rabbits or goats are most common and some prefer to use more than one species of animals to increase the diversity of the antibody population with the aim of obtaining better coverage.…”

Section: Development Of Immunoassays For Hcp Quantitationmentioning

confidence: 99%

“…Currently, there are several commercial HCP detection kits available for industrial use that biotechnology companies continue to evaluate and to compare with their own, as well as develop their own customized kits (Champion et al, 2001;Eaton, 1995;Hoffman, 2000). For commercial ELISAs, the main advantage is that no time and resources are necessary for antibody generation, method development, and quality control.…”

Section: Generic Versus Process-specific Hcp Assaysmentioning

confidence: 99%

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

Wang

Hunter

Mozier

2009

Biotech & Bioengineering

202

184

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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 present in small quantities (parts per million expressed as nanograms per milligrams of the intended recombinant protein) much effort and cost is expended by industry to remove them. The purpose of this review is to summarize what is of relevance in regards to the biology, the impact of genomics and proteomics on HCP evaluation, the regulatory expectations, analytical approaches, and various methodologies to remove HCPs with bioprocessing. Historical data, bioinformatics approaches and industrial case study examples are provided. Finally, a proposal for a risk assessment tool is provided which brings these facets together and proposes a means for manufacturers to classify and organize a control strategy leading to meaningful product specifications.

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“…Due to their nonhuman origin and thus potential immunogenic nature, HCPs can pose significant safety risk for patients and are part of process-related impurities that need to be controlled during bioprocess development [6][7][8]. Since after the purification steps, the residual HCPs amount in final drug substance is often very low in the parts per million (ppm) level, a highly specific, highly sensitive, and quantitative assay is desired to ensure their adequate removal and patient safety [9,10]. Due to its high specificity and sensivity, enzyme-linked immunosorbent assay (ELISA) is the most commonly accepted method by regulators for HCPs quantification [10].…”

Section: Introductionmentioning

confidence: 99%

The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics

Driscoll¹

2013

J Bioproces Biotechniq

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Biologics manufacturing requires the clearance of Host Cell Proteins (HCPs) from recombinant therapeutic protein to acceptable low levels to ensure product purity and patient safety. To ensure adequate removal, a highly sensitive method, commonly in the form of Enzyme-Linked Immunosorbent Assay (ELISA), is necessary to quantify the HCPs amount in process intermediates and drug substance. We report the development of a chemiluminescent detection based ELISA (luminescent ELISA) in lieu of previously used colorimetric method (colorimetric ELISA) to improve assay sensitivity for the quantification of Chinese Hamster Ovary (CHO) HCPs in a monoclonal antibody product (mAb-A). For luminescent ELISA, Pierce Supersignal ELISA Femto was chosen as the substrate to replace colorimetric substrate TMB. The assay performance of luminescent and colorimetric ELISA was directly compared side-by-side. Our data show that luminescent ELISA has better signal/background ratio, broader linear range over logarithmic scales, and better linearity within the same linear range than colorimetric ELISA. Luminescent ELISA also demonstrates better lowend linearity, greater accuracy and precision. In addition, the Limit of Detection (LOD) and Limit of Quantification (LOQ) are significantly improved with luminescent ELISA as compared to colorimetric ELISA. In summary, luminescent ELISA is a more sensitive method and demonstrates superiority over colorimetric method for CHO HCP quantification.

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Host cell contaminant protein assay development for recombinant biopharmaceuticals

Cited by 89 publications

References 37 publications

Proteomic Profiling of Secreted Proteins from CHO Cells Using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Proteomic Profiling of Secreted Proteins from CHO Cells Using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry

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

The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics

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