Novel Strategy Using Tryptic Peptide Immunoaffinity-Based LC–MS/MS to Quantify Denosumab in Monkey Serum

Wang, Qing; Han, Jiaguang; Sha, Chunjie; Liang, Yuanyingzhu; Sun, Yiying; Shao, Xia; Sun, Ji-Ye; Liu, Wanhui

doi:10.4155/bio-2017-0106

Cited by 9 publications

(2 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To ensure the sensitivity of quantitative analysis, the predicted tryptic peptides with 25 amino acids or more were not considered in the present study because of the difficulties in LC–MS/MS detection. The appropriate surrogate peptide should possess the following: it should be unique (possess a unique sequence) and representative, as well as possess several characteristics, for example, being free from interference from other proteins, added reagents, and/or other endogenous materials from the sample matrix using the detection method; it should consist of small peptides with 7–20 amino acids, because fewer amino acids would be disturbed by too many other kinds of interference peaks and more amino acids would be beyond the detection limit; it should avoid amino acids that are susceptible to modification during the digestion process and overall bioanalytical procedure, including methionine, asparagine, tryptophan, and glutamine; and it must produce an excellent chromatographic signal generated by selected reaction monitoring transition for the benefit of sufficient sensitivity (Wang et al, 2017).…”

Section: Resultsmentioning

confidence: 99%

Method development and validation of LC–MS/MS‐based assay for the simultaneous quantitation of trastuzumab and pertuzumab in cynomolgus monkey serum and its application in pharmacokinetic study

Gui

Dong

et al. 2020

Biomedical Chromatography

View full text Add to dashboard Cite

We present a simple and robust LC–MS/MS assay for the simultaneous quantitation of an antibody cocktail of trastuzumab and pertuzumab in monkey serum. The LC–MS/MS method saved costs, decreased the analysis time, and reduced quantitative times relative to the traditional ligand‐binding assays. The serum samples were digested with trypsin at 50°C for 60 min after methanol precipitation, ammonium bicarbonate denaturation, dithiothreitol reduction, and iodoacetamide alkylation. The tryptic peptides were chromatographically separated using a C18 column (2.1 × 50 mm, 2.6 μm) with mobile phases of 0.1% formic acid in water and acetonitrile. The other monoclonal antibody, infliximab, was used as internal standards to minimize the variability during sample processing and detection. A unique peptide for each monoclonal antibody was simultaneously quantified using LC–MS/MS in the multiple reaction monitoring mode. Calibration curves were linear from 2.0 to 400 μg/mL. The intra‐ and inter‐assay precision (%CV) was within 8.9 and 7.4% (except 10.4 and 15.1% for lower limit of quantitation), respectively, and the accuracy (%Dev) was within ±13.1%. The other validation parameters were evaluated, and all results met the acceptance criteria of the international guiding principles. Finally, the method was successfully applied to a pharmacokinetics study after a single‐dose intravenous drip administration to cynomolgus monkeys.

show abstract

Section: Resultsmentioning

confidence: 99%

Method development and validation of LC–MS/MS‐based assay for the simultaneous quantitation of trastuzumab and pertuzumab in cynomolgus monkey serum and its application in pharmacokinetic study

Gui

Dong

et al. 2020

Biomedical Chromatography

View full text Add to dashboard Cite

show abstract

“…A more selective alternative is affinity capture using an antibody directed to the constant (Fc) part of human IgG to selectively isolate a variety of human or humanized mAbs from animal matrices [25,26]. Like in LBAs, ultimate selectivity is obtained with a capturing reagent that is specifically directed to the analyte of interest, for example, an anti-idiotypic antibody [27] or the pharmacological target [28]. In this way, sensitivity down to the pg/ml level can typically be reached.…”

Section: Affinity Capturementioning

confidence: 99%

Protein Quantification by LC–MS: A Decade of Progress Through the Pages of Bioanalysis

Merbel

2019

Bioanalysis

View full text Add to dashboard Cite

Over the past 10 years, there has been a remarkable increase in the use of LC–MS for the quantitative determination of proteins, and this technique can now be considered an established bioanalytical platform for the quantification of macromolecular drugs and biomarkers, next to the traditional ligand-binding assays. Many researchers have contributed to the field and helped improve both the technical possibilities of LC–MS-based workflows and our understanding of the meaning of the results that are obtained. As a tribute to Bioanalysis, which has published many important contributions, this report gives a high-level overview of the most important trends in the field of protein LC–MS, as published in this journal since its inauguration a decade ago. It describes the major technical developments with regard to sample handling, separation and MS detection of both digested and intact protein analysis. In addition, the relevance of the complex structure and in vivo behavior of proteins is discussed and the effect of protein–protein interactions, biotransformation and the occurrence of isoforms on the analytical result is addressed.

show abstract

Analytical Challenges Assessing Protein Aggregation and Fragmentation Under Physiologic Conditions

Schuster

Mahler²,

Joerg³

et al. 2021

Journal of Pharmaceutical Sciences

View full text Add to dashboard Cite

Novel Strategy Using Tryptic Peptide Immunoaffinity-Based LC–MS/MS to Quantify Denosumab in Monkey Serum

Abstract: This approach accelerated the quantification, reduced the costs and provided an alternative in case of lacking the special antigen to denosumab or a RANKL-biotinylated reagent.

Cited by 9 publications

References 29 publications

Method development and validation of LC–MS/MS‐based assay for the simultaneous quantitation of trastuzumab and pertuzumab in cynomolgus monkey serum and its application in pharmacokinetic study

Method development and validation of LC–MS/MS‐based assay for the simultaneous quantitation of trastuzumab and pertuzumab in cynomolgus monkey serum and its application in pharmacokinetic study

Protein Quantification by LC–MS: A Decade of Progress Through the Pages of Bioanalysis

Analytical Challenges Assessing Protein Aggregation and Fragmentation Under Physiologic Conditions

Contact Info

Product

Resources

About