Quantitation of DNA and protein impurities in biopharmaceuticals

Briggs, J.; Panfili, Peter R.

doi:10.1021/ac00009a003

Cited by 78 publications

(32 citation statements)

References 29 publications

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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%

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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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Section: Analytical Technology Overviewmentioning

confidence: 99%

Section: Development Of Immunoassays For Hcp Quantitationmentioning

confidence: 99%

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

Wang

Hunter

Mozier

2009

Biotech & Bioengineering

202

184

View full text Add to dashboard Cite

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“…Concerns regarding contamination by genomic DNA are based on the possibility of an oncogene being encoded by those fragments and its possible activation or deactivation once in the recipient cells [20,21]. Although the risk is very low ± less than 10 ±9 tumor formation frequency was reported after the in vivo administration of a 1 ng genomic DNA dose to cells containing one hundred copies of oncogenic DNA [21] ± limits are imposed by regulatory agencies with the sole concern of patient safety [19,22].…”

Section: Host Nucleic Acidsmentioning

confidence: 99%

Chromatographic Approaches in the Purification of Plasmid DNA for Therapy and Vaccination

Ferreira

2005

Chem Eng & Technol

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The need for large scale plasmid DNA purification processes has increased in the last few years due to the rapid evolution of gene therapy and DNA vaccines. Chromatographic purification of plasmid DNA has specific problems which are mostly related to the structural nature of plasmid DNA, the resemblance of plasmid DNA to some impurities, and the lack of capacity and selectivity of the traditional bead adsorbents. In this paper, plasmid DNA purification using chromatographic processes is reviewed. Initially the most relevant structural and chemical properties of supercoiled plasmid DNA and related impurities are described, and the purification objectives are specified. The most significant properties are highlighted and their possible exploitation for plasmid DNA adsorption is described. After a brief description of currently used chromatographic purification strategies, novel approaches centered on the use and development of new stationary phases with higher capacity utilization are described.

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“…However, the harvest also contains significant amounts of proteins originated from the host, namely host cell proteins (HCPs), which are either secreted during fermentation or released into culture fluid as a result of cell lysis. 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].…”

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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Quantitation of DNA and protein impurities in biopharmaceuticals

Cited by 78 publications

References 29 publications

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

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

Chromatographic Approaches in the Purification of Plasmid DNA for Therapy and Vaccination

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

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