Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination
Host cell proteins (HCPs) must be sufficiently cleared from recombinant biopharmaceuticals during the downstream process (DSP) to ensure product quality, purity, and patient safety. For monitoring of HCP clearance, the typical method chosen is an enzyme-linked immunosorbent assay (ELISA) using polyc...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2021-01-01
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| Series: | mAbs |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2021.1955432 |
| _version_ | 1798040803563536384 |
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| author | Daniel M. Waldera-Lupa Yvonne Jasper Pia Köhne Ronja Schwichtenhövel Heiner Falkenberg Thomas Flad Peter Happersberger Bernd Reisinger Alireza Dehghani Roland Moussa Thomas Waerner |
| author_facet | Daniel M. Waldera-Lupa Yvonne Jasper Pia Köhne Ronja Schwichtenhövel Heiner Falkenberg Thomas Flad Peter Happersberger Bernd Reisinger Alireza Dehghani Roland Moussa Thomas Waerner |
| author_sort | Daniel M. Waldera-Lupa |
| collection | DOAJ |
| description | Host cell proteins (HCPs) must be sufficiently cleared from recombinant biopharmaceuticals during the downstream process (DSP) to ensure product quality, purity, and patient safety. For monitoring of HCP clearance, the typical method chosen is an enzyme-linked immunosorbent assay (ELISA) using polyclonal anti-HCP antibodies obtained from an immunization campaign. This polyclonal reagent is a critical factor for functionality and confidence of the ELISA. Therefore, it is important to ensure that the pool of ELISA antibodies covers a broad spectrum of the HCPs that potentially could persist in the final drug substance. Typically, coverage is determined by gel-based approaches. Here, we present a quantitative proteomics approach combined with purification of HCPs by immunoaffinity chromatography (qIAC-MS) for assessment of ELISA coverage. The cell culture fluid (CCF) of a mock fermentation and a recombinant monoclonal antibody product were characterized in detail to investigate whether the HCPs used for immunization of animals accurately represent HCPs that are relevant to the process. Using the qIAC-MS approach, the ELISA antibody coverage was determined for mock fermentation and product CCF, as well as several different DSP intermediates. Here, the use of different controls facilitated the identification and quantification of HCPs present in the polyclonal reagent and those that nonspecifically bound to IAC material. This study successfully demonstrates that the described qIAC-MS approach is not only a suitable orthogonal method to commonly used 2D SDS-PAGE-based analysis for evaluating ELISA antibody coverage, but that it further identifies HCPs covered as well as missed by the ELISA, enabling an improved risk assessment of HCP ELISA. |
| first_indexed | 2024-04-11T22:12:42Z |
| format | Article |
| id | doaj.art-8c24c9a2e4674e379ac17b043d12ac94 |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2024-04-11T22:12:42Z |
| publishDate | 2021-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-8c24c9a2e4674e379ac17b043d12ac942022-12-22T04:00:31ZengTaylor & Francis GroupmAbs1942-08621942-08702021-01-0113110.1080/19420862.2021.1955432Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determinationDaniel M. Waldera-Lupa0Yvonne Jasper1Pia Köhne2Ronja Schwichtenhövel3Heiner Falkenberg4Thomas Flad5Peter Happersberger6Bernd Reisinger7Alireza Dehghani8Roland Moussa9Thomas Waerner10Bioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyBioanalytics, Protagen Protein Services GmbH, Dortmund, GermanyAnalytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyHost cell proteins (HCPs) must be sufficiently cleared from recombinant biopharmaceuticals during the downstream process (DSP) to ensure product quality, purity, and patient safety. For monitoring of HCP clearance, the typical method chosen is an enzyme-linked immunosorbent assay (ELISA) using polyclonal anti-HCP antibodies obtained from an immunization campaign. This polyclonal reagent is a critical factor for functionality and confidence of the ELISA. Therefore, it is important to ensure that the pool of ELISA antibodies covers a broad spectrum of the HCPs that potentially could persist in the final drug substance. Typically, coverage is determined by gel-based approaches. Here, we present a quantitative proteomics approach combined with purification of HCPs by immunoaffinity chromatography (qIAC-MS) for assessment of ELISA coverage. The cell culture fluid (CCF) of a mock fermentation and a recombinant monoclonal antibody product were characterized in detail to investigate whether the HCPs used for immunization of animals accurately represent HCPs that are relevant to the process. Using the qIAC-MS approach, the ELISA antibody coverage was determined for mock fermentation and product CCF, as well as several different DSP intermediates. Here, the use of different controls facilitated the identification and quantification of HCPs present in the polyclonal reagent and those that nonspecifically bound to IAC material. This study successfully demonstrates that the described qIAC-MS approach is not only a suitable orthogonal method to commonly used 2D SDS-PAGE-based analysis for evaluating ELISA antibody coverage, but that it further identifies HCPs covered as well as missed by the ELISA, enabling an improved risk assessment of HCP ELISA.https://www.tandfonline.com/doi/10.1080/19420862.2021.1955432Host cell proteinmass spectrometryproteomicsbiopharmaceuticalsELISA coverageaffinity purification |
| spellingShingle | Daniel M. Waldera-Lupa Yvonne Jasper Pia Köhne Ronja Schwichtenhövel Heiner Falkenberg Thomas Flad Peter Happersberger Bernd Reisinger Alireza Dehghani Roland Moussa Thomas Waerner Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination mAbs Host cell protein mass spectrometry proteomics biopharmaceuticals ELISA coverage affinity purification |
| title | Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination |
| title_full | Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination |
| title_fullStr | Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination |
| title_full_unstemmed | Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination |
| title_short | Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage determination |
| title_sort | host cell protein detection gap risk mitigation quantitative iac ms for elisa antibody reagent coverage determination |
| topic | Host cell protein mass spectrometry proteomics biopharmaceuticals ELISA coverage affinity purification |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2021.1955432 |
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