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Bioanalysis. 2015;7(10):1237-51. doi: 10.4155/bio.15.56. Epub 2015 Apr 21.

Absolute and multiplex quantification of antibodies in serum using PSAQ™ standards and LC-MS/MS.

Author information

1
1PROMISE Advanced Proteomics, Grenoble, F-38040, France.
2
2Centre d'Immunologie Pierre Fabre (CIPF), 5 Av. Napoléon III, BP 60497, 74164 Saint-Julien-en-Genevois, France.
3
3Centre d'Etudes Pré-Cliniques Pierre Fabre, Campans, France.
4
4PX-Therapeutics, Grenoble, F-38040, France.
5
5Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France.
6
6CEA, iRTSV-BGE, F-38000 Grenoble, France.
7
7INSERM, BGE, F-38000 Grenoble, France.

Abstract

BACKGROUND:

In preclinical studies, monoclonal antibodies (mAbs) are traditionally assayed by ligand-binding-assays. Recently, quantitative liquid chromatography mass spectrometry (MS)-based assays have emerged which circumvent a number of challenges. These assays may also be multiplex, making them potentially compatible with pharmacokinetic assays for combined antibody therapies.

MATERIALS & METHODS:

We combined a quantitative MS-based approach with the protein standard for absolute quantification (PSAQ™) strategy to simultaneously quantify three mAb variants presenting minor sequence differences. Stable isotopically labeled mAbs were produced and used as quantification standards. Titration curves were performed to assess the analytical performances of the method. LC-MS/MS and ELISA data were cross-compared.

RESULTS:

The approach presented provides similar accuracy and precision than ELISA, while being multiplex and faster to develop. It has applications at all stages of the pharmaceutical development.

PMID:
25898209
DOI:
10.4155/bio.15.56
[Indexed for MEDLINE]

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