LC-MS bioanalysis of intact proteins and peptides.
Lijuan KangNaidong WengWenying JianPublished in: Biomedical chromatography : BMC (2019)
Bioanalysis assays that reliably quantify biotherapeutics and biomarkers in biological samples play pivotal roles in drug discovery and development. Liquid chromatography coupled with mass spectrometry (LC-MS), owing to its superior specificity, faster method development and multiplex capability, has evolved as one of the most important platforms for bioanalysis of biotherapeutics, particularly new scaffolds such as half-life extension platforms for proteins and peptides, as well as antibody drug conjugates. Intact LC-MS analysis is orthogonal to bottom-up surrogate peptide approach by providing whole molecule quantitation and high-level sequence and structure information. Here we review the latest development in LC-MS bioanalysis of intact proteins and peptides by summarizing recent publications and discussing the important topics such as the comparison between top-down intact analysis and bottom-up surrogate peptide approach, as well as simultaneous quantitation and catabolite identification. Key bioanalytical issues around intact protein bioanalysis such as sensitivity, data processing strategies, specificity, sample preparation and LC condition are elaborated. For peptides, topics including quantitation of intact peptide vs. digested surrogate peptide, metabolites, sensitivity, LC condition, assay performance, internal standard and sample preparation are discussed.
Keyphrases
- liquid chromatography
- mass spectrometry
- tandem mass spectrometry
- high resolution mass spectrometry
- ms ms
- simultaneous determination
- high performance liquid chromatography
- drug discovery
- amino acid
- liquid chromatography tandem mass spectrometry
- solid phase extraction
- high throughput
- gas chromatography
- capillary electrophoresis
- high resolution
- drug delivery
- social media
- cancer therapy
- structural basis
- small molecule
- sewage sludge
- bioinformatics analysis
- data analysis