Recommendations for Validation of LC-MS/MS Bioanalytical Methods for Protein Biotherapeutics
Open accessAbstract
This paper represents the consensus views of a cross-section of companies and organizations from the USA and Canada regarding the validation and application of liquid chromatography tandem mass spectrometry (LC-MS/MS) methods for bioanalysis of protein biotherapeutics in regulated studies. It was prepared under the auspices of the AAPS Bioanalytical Focus Group's Protein LC-MS Bioanalysis Subteam and is intended to serve as a guide to drive harmonization of best practices within the bioanalytical community and provide regulators with an overview of current industry thinking on applying LC-MS/MS technology for protein bioanalysis. For simplicity, the scope was limited to the most common current approach in which the protein is indirectly quantified using LC-MS/MS measurement of one or more of its surrogate peptide(s) produced by proteolytic digestion. Within this context, we considered a range of sample preparation approaches from simple in-matrix protein denaturation and digestion to complex procedures involving affinity capture enrichment. Consideration was given to the method validation experiments normally associated with traditional LC-MS/MS and ligand-binding assays. Our collective experience, thus far, is that LC-MS/MS methods for protein bioanalysis require different development and validation considerations than those used for small molecules. The method development and validation plans need to be tailored to the particular assay format being established, taking into account a number of important factors: the intended use of the assay, the test species or study population, the characteristics of the protein biotherapeutic and its similarity to endogenous proteins, potential interferences, as well as the nature, quality, and availability of reference and internal standard materials.
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LC-MS/MS methods also have problems with sample matrix interference.
“Such methods also have problems with sample matrix interference 124.”
From Measuring Oxytocin and Vasopressin: Bioassays, Immunoassays and Random Numbers (2016) · cites this paper as [124]
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Measuring Oxytocin and Vasopressin: Bioassays, Immunoassays and Random Numbers
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“Such methods also have problems with sample matrix interference (124).”
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- Authors
- Jenkins, R., Duggan, J. X., Aubry, A.-F., Zeng, J., Lee, J. W., Cojocaru, L., Dufield, D., Garofolo, F., Kaur, S., Schultz, G. A., Xu, K., Yang, Z., Yu, J., Zhang, Y. J., & Vazvaei, F.
- Affiliations
- Pharmaceutical Product Development (United States) (US)
- Boehringer Ingelheim (United States) (US)
- Bristol-Myers Squibb (United States) (US)
- Pfizer (United States) (US)
- Genentech (US)
- IQVIA (United Kingdom) (GB)
- Corresponding author
- Vazvaei, Faye
- Venue
- The AAPS Journal
- Published
- 2014-11-13
- Volume
- 17
- Issue
- 1
- Pages
- 1-16
- ISSN
- 1550-7416 (ISSN-L)
- PMC
- PMC4287296
- OpenAlex
- W1964595219
- Cited by
- 181
- References
- 27
- Citations by year
- 2026: 9 · 2025: 11 · 2024: 15 · 2023: 9 · 2022: 9 · 2021: 14 · 2020: 18 · 2019: 18 · 2018: 26 · 2017: 23 · 2016: 22 · 2015: 7
- FWCI
- 7.92
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- The AAPS Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Bristol-Myers Squibb
- Eli Lilly and Company
- Pfizer
- GlaxoSmithKline
- Biogen
- Genentech
- Novartis Institutes for BioMedical Research
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Biosimilars and Bioanalytical Methods
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- method validation, surrogate peptides, proteolytic digestion, sample preparation, method development, reference materials, internal standard, regulatory harmonization
- MeSH terms
- Animals, Canada, Chromatography, Liquid, Humans, Proteins, United States, Tandem Mass Spectrometry, Validation Studies as Topic
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