Ultra‐performance liquid chromatography/multiple reaction monitoring mass spectrometry quantification of trastuzumab in human serum by selective monitoring of a specific peptide marker from the antibody complementarity‐determining regions

Russo, Rosita; Rega, Camilla; Caporale, Andrea; Tonon, Giancarlo; Scaramuzza, Silvia; Selis, Fabio; Ruvo, Menotti

doi:10.1002/rcm.7898

Cited by 15 publications

(11 citation statements)

References 21 publications

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“…Recently, it has been reported that the expression of an scFv against HER2 derived from Trastuzumab (drug bank number DB00072) in SHuffle at 30 • C resulted in enhanced solubility and a higher expression level of the molecule as compared to its expression in BL21 (DE3) at 37 • C [109][110][111][112].…”

Section: Cytoplasmic Expressionmentioning

confidence: 99%

Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Sandomenico

Sivaccumar

Ruvo

2020

IJMS

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Antibodies and antibody-derived molecules are continuously developed as both therapeutic agents and key reagents for advanced diagnostic investigations. Their application in these fields has indeed greatly expanded the demand of these molecules and the need for their production in high yield and purity. While full-length antibodies require mammalian expression systems due to the occurrence of functionally and structurally important glycosylations, most antibody fragments and antibody-like molecules are non-glycosylated and can be more conveniently prepared in E. coli-based expression platforms. We propose here an updated survey of the most effective and appropriate methods of preparation of antibody fragments that exploit E. coli as an expression background and review the pros and cons of the different platforms available today. Around 250 references accompany and complete the review together with some lists of the most important new antibody-like molecules that are on the market or are being developed as new biotherapeutics or diagnostic agents.

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Section: Cytoplasmic Expressionmentioning

confidence: 99%

Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Sandomenico

Sivaccumar

Ruvo

2020

IJMS

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“…Essential for minimizing technical variations during sample preparation is the addition of an internal standard (IS), displaying similar or identical physical and chemical features, which allows monitoring of the full sample preparation workflow.- 10,12,14 LC-MS/MS methods using multiple reaction monitoring (MRM) for quantification of only trastuzumab in human serum have been described previously, including a method employing two surrogate peptides reaching a sensitivity of 5 μg/mL, and another reaching a sensitivity of 20 ng/mL with the latter using high resolution MS detection. [15][16][17][18] To facilitate quantification of co-administered mAbs, multiplex LC-MS/MS methods have been developed. However, until recently these assays always required specific reagents for affinity purification or specific stable isotope labeled (SIL) ISs.…”

Section: Introductionmentioning

confidence: 99%

Development and validation of an LC-MS/MS method for simultaneous quantification of co-administered trastuzumab and pertuzumab

Schokker

Bonardi

Molenaar

et al. 2020

mAbs

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Given the increasing use of combination therapy with multiple monoclonal antibodies (mAbs), there is a clinical need for multiplexing assays. For the frequently co-administered anti-human epidermal growth factor receptor 2 (HER2) mAbs trastuzumab and pertuzumab, we developed a high-throughput and robust hybrid ligand-binding liquid chromatography-mass spectrometry (LC-MS)/MS quantitative assay. Nanomolar concentrations of trastuzumab and pertuzumab were determined in 10 µL serum samples after extraction by affinity purification through protein A beads, followed by on-bead reduction, alkylation, and trypsin digestion. After electrospray ionization, quantification was obtained by multiple reaction monitoring LC-MS/MS using SILuMab as an internal standard. The method was validated according to the current guidelines from the US Food and Drug Administration and the European Medicines Agency. Assay linearity was established in the ranges 0.250-250 μg/mL for trastuzumab and 0.500-500 μg/mL for pertuzumab. The method was accurate and selective for the simultaneous determination of trastuzumab and pertuzumab in clinical samples, thereby overcoming the limitation of ligand binding assays that cannot quantify mAbs targeting the same receptor. Furthermore, this method requires a small blood volume, which reduces blood collection time and stress for patients. The assay robustness was verified in a clinical trial where trastuzumab and pertuzumab concentrations were determined in 670 serum samples. As we used commercially available reagents and standards, the described generic bioanalytical strategy can easily be adapted to multiplex quantifications of other mAb combinations in non-clinical and clinical samples.

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“…Wang et al reported that two organosulfur compounds without matched library data were inferred according to the tandem mass spectra obtained using GC/Q‐TOF . Compounds in unknown complexes can be identified via selective/multiple reaction monitoring (SRM/MRM) . SRM/MRM methods run by GC/Q‐TOF MS were shown to eliminate most non‐target detection and improve specificity and sensitivity …”

Section: Introductionmentioning

confidence: 99%

“…27 Compounds in unknown complexes can be identified via selective/multiple reaction monitoring (SRM/MRM). 31,32 SRM/MRM methods run by GC/Q-TOF MS were shown to eliminate most non-target detection and improve specificity and sensitivity. 33,34 The identification of compounds in coal using GC/MS can be disturbed by co-eluted species, 35,36 which hamper the monitoring of coal conversions like hydrodeoxygenation, hydrodenitrogenation and hydrodesulfurization.…”

mentioning

confidence: 99%

Evaluation of coal‐related model compounds using tandem mass spectrometry

Dong

Fan

et al. 2018

Rapid Comm Mass Spectrometry

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Ultra‐performance liquid chromatography/multiple reaction monitoring mass spectrometry quantification of trastuzumab in human serum by selective monitoring of a specific peptide marker from the antibody complementarity‐determining regions

Abstract: The strategy used to set up the UPLC/MRM MS methodology based on monitoring specific peptide markers within CDRs can be potentially applied to the detection and quantification of other humanized or human mAbs in biological fluids. Copyright © 2017 John Wiley & Sons, Ltd.

Cited by 15 publications

References 21 publications

Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Development and validation of an LC-MS/MS method for simultaneous quantification of co-administered trastuzumab and pertuzumab

Evaluation of coal‐related model compounds using tandem mass spectrometry

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