Immuno‐mass spectrometry assay of EPI‐HNE4, a recombinant protein inhibitor of human elastase

Dubois, Mathieu; Bécher, François; Herbet, Amaury; Ezan, Éric

doi:10.1002/rcm.2844

Cited by 38 publications

(37 citation statements)

References 21 publications

(28 reference statements)

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“…Although this technique requires specialized antibodies, it provides sufficient purification to achieve quantification of low abundance proteins from plasma [24,[28][29][30][31]. Low recoveries and cross reactivities are some of the issues seen during immunocapture enrichment [32,33]. Immunoaffinity isolations may be carried out with single or multiple antibodies depending on the availability of analyte-specific antibodies and the desired detection limits.…”

Section: Immunoaffinity Enrichmentmentioning

confidence: 99%

Internal Standards for Absolute Quantification of Large Molecules (Proteins) from Biological Matrices by LC-MS/MS

Faria¹,

Halquist²

2018

Calibration and Validation of Analytical Methods - A Sampling of Current Approaches

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Internal standardization plays a critical role in the performance of a bioanalytical method. There has been a tremendous increase in the popularity of using liquid chromatography tandem mass spectrometry (LC-MS/MS) methods for quantitative bioanalysis of protein molecules. Protein, being too large to be directly analyzed by LC-MS/MS, is proteolyzed and a characteristic peptide is used as a surrogate analyte for quantification. Internal standardization in small molecules' analysis is straightforward, i.e., either a stable labeled isotope (SIL) form of the analyte or a structural analogue is used. As protein quantification involves protein digestion to yield peptides, there are more options for internal standardization. Currently, internal standard selection is based on the availability of the internal standards and the sample preparation workflow. A SIL-form of the analyte protein is the ideal internal standard. However, its use is limited due to cost and commercial availability. Alternatively, a SIL form the surrogate peptide analyte or a cleavable SIL-peptide can be used as an IS. For preclinical bioanalysis of humanized IgG antibody-based drugs, a universal SIL analogue protein has been effectively used as an internal standard. This chapter focuses on internal standardization for the quantitative analysis of proteins, such as biotherapeutics and biomarkers, using LC-MS/MS.

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Section: Immunoaffinity Enrichmentmentioning

confidence: 99%

Internal Standards for Absolute Quantification of Large Molecules (Proteins) from Biological Matrices by LC-MS/MS

Faria¹,

Halquist²

2018

Calibration and Validation of Analytical Methods - A Sampling of Current Approaches

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“…Liquid chromatography-tandem mass spectrometric (LC-MS/MS) methods coupled with immunoaffinity sample enrichment have gained interest due to the potential advantages of additional specificity, reduced need for specific reagents, and faster method development time for the analysis of biologics (1)(2)(3)(4)(5)(6). We have recently reported the approach of using a common immunoaffinity capture and a general stable isotope-labeled whole-antibody internal standard (SIL-mAb IS) incorporating an essential SIL-amino acid.…”

Section: Introductionmentioning

confidence: 99%

“…There are two objectives of this paper: (1) to present the method development and qualification of a multiplex LC-MS/ MS method for four mAbs and (2) to demonstrate the applicability of the multiplex LC-MS/MS method in the analysis of samples from cassette-dosed rats and compare the results to those of discrete-dosed rats. The potential effects of immunogenicity and target-binding on the PK results from cassette-dosing of mAbs were discussed.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Analysis of Multiple Monoclonal Antibody Biotherapeutics by LC-MS/MS Method in Rat Plasma Following Cassette-Dosing

Ortiz

Tran

et al. 2012

AAPS J

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Abstract. We have recently developed a general liquid chromatography-tandem mass spectrometric (LC-MS/MS) method using a stable isotope-labeled (SIL) monoclonal antibody (mAb) as an internal standard (IS) for single-analyte quantification of mAb (Li et al. Anal Chem 84(3):1267-1273, 2012. The method offers an advantage over ligand binding assay in reducing the time and resources needed for bioanalytical support in preclinical stages of drug development. In this paper, we report another marked increase in assay efficiency for multi-analyte bioanalysis using unique surrogate peptides for each analyte and the strategic choice of the SIL-IS peptide. The method was qualified for the simultaneous determinations of four mAbs in rat plasma and applied to samples from discrete-and cassette-dosed rats. The pharmacokinetic parameters of the four mAbs of cassette dosing were comparable to those of discrete dosing and of enzyme-linked immunosorbent assay results. Although there may be limitations and special considerations for cassette-dosing of biologics, these results demonstrate the robust performance of the multi-analyte LC-MS/MS method allowing cassette-dosing that would ultimately reduce animal use and improve efficiency.KEY WORDS: cassette dosing of biologics; immunoaffinity-mass spectrometry; ligand binding assay; monoclonal antibody biotherapeutics; multi-analyte LC-MS/MS.

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“…Biotherapeutics or associated surrogate peptides as well as any important catabolites could be specifically quantified by MS. In fact, several researchers have published studies with this ultimate goal in mind (Dubois et al, 2007(Dubois et al, , 2008Heudi et al, 2008;Ezan et al, 2009;Liu et al, 2011b;Bronsema et al, 2012;Li et al, 2012). Historically, there have been several drawbacks associated with quantitative MS methods compared with LBA, including the following: 1) MS has been generally less sensitive, 2) large protein analytes have suffered from low throughput, and 3) quantitative MS methods for macromolecules are much more nascent compared with established antibody-based methodologies.…”

Section: Impact Of Protein Biotherapeutic Biotransformation On Bioanamentioning

confidence: 99%

Biotransformation and In Vivo Stability of Protein Biotherapeutics: Impact on Candidate Selection and Pharmacokinetic Profiling

Hall

2014

Drug Metab Dispos

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Historically, since the metabolism of administered peptide/protein drugs ("biotherapeutics") has been expected to undergo predictable pathways similar to endogenous proteins, comprehensive biotherapeutic metabolism studies have not been widely reported in the literature. However, since biotherapeutics have rapidly evolved into an impressive array of eclectic modalities, there has been a shift toward understanding the impact of metabolism on biotherapeutic development. For biotherapeutics containing non-native chemical linkers and other moieties besides natural amino acids, metabolism studies are critical as these moieties may impart undesired toxicology. For biotherapeutics that are composed solely of natural amino acids, where end-stage peptide and amino acid catabolites do not generally pose toxicity concerns, the understanding of biotherapeutic biotransformation, defined as in vivo modifications such as peripherally generated intermediate circulating catabolites prior to end-stage degradation or elimination, may impact in vivo stability and potency/clearance. As of yet, there are no harmonized methodologies for understanding biotherapeutic biotransformation and its impact on drug development, nor is there clear guidance from regulatory agencies on how and when these studies should be conducted. This review provides an update on biotherapeutic biotransformation studies and an overview of lessons learned, tools that have been developed, and suggestions of approaches to address issues. Biotherapeutic biotransformation studies, especially for certain modalities, should be implemented at an early stage of development to 1) understand the impact on potency/clearance, 2) select the most stable candidates or direct protein re-engineering efforts, and 3) select the best bioanalytical technique(s) for proper drug quantification and subsequent pharmacokinetic profiling and exposure/response assessment.

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Immuno‐mass spectrometry assay of EPI‐HNE4, a recombinant protein inhibitor of human elastase

Cited by 38 publications

References 21 publications

Internal Standards for Absolute Quantification of Large Molecules (Proteins) from Biological Matrices by LC-MS/MS

Internal Standards for Absolute Quantification of Large Molecules (Proteins) from Biological Matrices by LC-MS/MS

Simultaneous Analysis of Multiple Monoclonal Antibody Biotherapeutics by LC-MS/MS Method in Rat Plasma Following Cassette-Dosing

Biotransformation and In Vivo Stability of Protein Biotherapeutics: Impact on Candidate Selection and Pharmacokinetic Profiling

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