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Statistical characterization of therapeutic protein modifications
- Source :
- Scientific Reports, Scientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
- Publication Year :
- 2017
- Publisher :
- Nature Publishing Group UK, 2017.
-
Abstract
- Peptide mapping with liquid chromatography–tandem mass spectrometry (LC-MS/MS) is an important analytical method for characterization of post-translational and chemical modifications in therapeutic proteins. Despite its importance, there is currently no consensus on the statistical analysis of the resulting data. In this manuscript, we distinguish three statistical goals for therapeutic protein characterization: (1) estimation of site occupancy of modifications in one condition, (2) detection of differential site occupancy between conditions, and (3) estimation of combined site occupancy across multiple modification sites. We propose an approach, which addresses these goals in terms of summarizing the quantitative information from the mass spectra, statistical modeling, and model-based analysis of LC-MS/MS data. We illustrate the approach using an LC-MS/MS experiment from an antibody-drug conjugate and its monoclonal antibody intermediate. The performance was compared to a ‘naïve’ data analysis approach, by using computer simulation, evaluation of differential site occupancy in positive and negative controls, and comparisons of estimated site occupancy with orthogonal experimental measurements of N-linked glycoforms and total oxidation. The results demonstrated the importance of replicated studies of protein characterization, and of appropriate statistical modeling, for reproducible, accurate and efficient site occupancy estimation and differential analysis.
- Subjects :
- 0301 basic medicine
medicine.drug_class
Computer science
Science
Computational biology
Biostatistics
Monoclonal antibody
Mass spectrometry
Tandem mass spectrometry
01 natural sciences
Peptide Mapping
Article
03 medical and health sciences
Tandem Mass Spectrometry
medicine
Technology, Pharmaceutical
Simulation
Chromatography, High Pressure Liquid
Biological Products
Multidisciplinary
Peptide mapping
010401 analytical chemistry
Therapeutic protein
Proteins
Statistical model
0104 chemical sciences
Characterization (materials science)
030104 developmental biology
Medicine
Protein Processing, Post-Translational
Subjects
Details
- Language :
- English
- ISSN :
- 20452322
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....5440a1281217169632d9cdfd807efd81