Improved identification of host cell proteins in a protein biopharmaceutical by LC–MS/MS using the ProteoMiner™ Enrichment Kit

Mörtstedt, Harriet; Makower, Åsa; Edlund, Per-Olof; Sjöberg, Katarina; Tjernberg, Agneta

doi:10.1016/j.jpba.2020.113256

Cited by 17 publications

(7 citation statements)

References 28 publications

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“…In recent years, multiple studies have employed DDA-based LC-MS/MS approaches to detect and quantify HCP impurities in several different mAb products. While these studies presented sensitive and reproducible orthogonal methods when compared to ELISA [ 2 ], most protocols require some form of sample pre-treatment to improve HCP detection [ 6 , 9 , 10 ]. However, the industry is looking for sample preparation and analysis procedures that are simple, reproducible, robust, and easy to implement in existing workflows.…”

Section: Resultsmentioning

confidence: 99%

“…One commonly applied approach to circumventing this problem is to resolve co-eluting peptides by pre-fractionation [ [6] , [7] , [8] ]. Another possibility is to reduce the dynamic range using molecular weight cutoff (MWCO) filters or by targeted depletion of the mAb prior to analysis [ 9 , 10 ]. Recently, alternative digestion approaches have been employed [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

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Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS

Strasser

Oliviero

Jakes

et al. 2021

Journal of Pharmaceutical Analysis

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Ensuring the removal of host cell proteins (HCPs) during downstream processing of recombinant proteins such as monoclonal antibodies (mAbs) remains a challenge. Since residual HCPs might affect product stability or safety, constant monitoring is required to demonstrate their removal to be below the regulatory accepted level of 100 ng/mg. The current standard analytical approach for this procedure is based on ELISA; however, this approach only measures the overall HCP content. Therefore, the use of orthogonal methods, such as liquid chromatography-mass spectrometry (LC-MS), has been established, as it facilitates the quantitation of total HCPs as well as the identification and quantitation of the individual HCPs present. In the present study, a workflow for HCP detection and quantitation using an automated magnetic bead-based sample preparation, in combination with a data-independent acquisition (DIA) LC-MS analysis, was established. Employing the same instrumental setup commonly used for peptide mapping analysis of mAbs allows for its quick and easy implementation into pre-existing workflows, avoiding the need for dedicated instrumentation or personnel. Thereby, quantitation of HCPs over a broad dynamic range was enabled to allow monitoring of problematic HCPs or to track changes upon altered bioprocessing conditions.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS

Strasser

Oliviero

Jakes

et al. 2021

Journal of Pharmaceutical Analysis

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“…Subsequently, impurities were successfully tracked from other biological preparations such as purified monoclonal antibodies [74,75]. This interesting technology was then forgotten for several years; however, its merit was recently ''rediscovered'' to track host cell proteins [76,77]. This field is not yet exploited as it should because antibodies against known host cell proteins are currently used.…”

Section: Evolution and Extension Of Cpll Technology To Various Fieldsmentioning

confidence: 99%

Combinatorial peptides: A library that continuously probes low‐abundance proteins

Boschetti¹,

Zilberstein

Righetti

2021

Electrophoresis

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After a decade of experimental applications, it is the objective of this review to make a point on combinatorial peptide ligand libraries dedicated to low‐abundance proteins from animals to plants and to microorganism proteomics. It is, thus, at the light of the recent technical developments and applications that we will examine the state of the art, its usage within the scientific community, and its openness to unexplored fields. The improvements of the methodology and its implementation in connection with analytical determinations of combinatorial peptide ligand library (CPLL)‐treated samples are extensively reviewed and commented upon. Relevant examples covering few critical aspects describe the performance of the technology. Finally, a reflection on the technological future is attempted in particular by involving new concepts adapted to the limited availability of certain biological samples.

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“…The method of the detection of HCP contained in the third volume (2020 edition) of China Pharmacopoeia 20 all use the double antibody sandwich ELISA method, 21 an international general method for the detection, which can be used to evaluate the ability of the biological agent production process to reduce or eliminate the non‐destination components. However, the results of commercial HCP detection kits 22,23 only represent the general content and proportion of universal HCPs, each project has its specific proportion of HCPs and the interference degree of each protein on the detection of HCPs is different, so it is necessary to establish a dedicated method for HCP detection. In this study, the process‐specific immunological detection method was adopted, by which the host cells were treated according to the product production and purification processes to obtain the process‐specific HCP which was used to prepare the standards and detection antibody.…”

Section: Introductionmentioning

confidence: 99%

“…However, the results of commercial HCP detection kits 22,23 only represent the general content and proportion of universal HCPs, each project has its specific proportion of HCPs and the interference degree of each protein on the detection of HCPs is different, so it is necessary to establish a dedicated method for HCP detection. In this study, the process-specific immunological detection method was adopted, by which the host cells were treated according to the product production and purification processes to obtain the processspecific HCP which was used to prepare the standards and detection antibody.…”

mentioning

confidence: 99%

Establishment and evaluation of detection methods for process‐specific residual host cell protein and residual host cell DNA in biological preparation

Tian,

Wang,

Shao

et al. 2024

Cell Biochemistry & Function

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To establish accurate detection methods of process‐specific Escherichia coli residual host cell protein (HCP) and residual host cell DNA (rcDNA) in recombinant biological preparations. Taking the purification process of GLP expressed by E. coli as a specific‐process model, the HCP of empty E. coli was intercepted to immunize mice and rabbits. Using IgG from immunized rabbits as the coating antibody and mouse immune serum as the second sandwich antibody, a process‐specific enzyme‐linked immunosorbent assay (ELISA) for E. coli HCP was established. Targeting the 16S gene of E. coli, ddPCR was used to obtain the absolute copies of rcDNA in samples. Non‐process‐specific commercial ELISA kit and the process‐specific ELISA established in this study were used to detect the HCP in GLP preparation. About 62% of HCPs, which should be process‐specific HCPs, could not be detected by the non‐process‐specific commercial ELISA kit. The sensitivity of established ELISA can reach 338 pg/mL. The rcDNA could be absolutely quantitated by ddPCR, for the copies of rcDNA in three multiple diluted samples showed a reduced gradient. While the copies of rcDNA in three multiple diluted samples could not be distinguished by the qPCR. Process‐specific ELISA has high sensitivity in detecting process‐specific E. coli HCP. The absolutely quantitative ddPCR has much higher accuracy than the relatively quantitative qPCR, it is a nucleic acid quantitative method that is expected to replace qPCR in the future.

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Improved identification of host cell proteins in a protein biopharmaceutical by LC–MS/MS using the ProteoMiner™ Enrichment Kit

Cited by 17 publications

References 28 publications

Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS

Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS

Combinatorial peptides: A library that continuously probes low‐abundance proteins

Establishment and evaluation of detection methods for process‐specific residual host cell protein and residual host cell DNA in biological preparation

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