Simultaneous extraction and analysis of multiple cystine-dense peptides by μSPE and microflow-MS/MS from plasma. (19th March 2019)

• Record Type: Journal Article
• Authors: Brusniak, Mi-Youn; Christianson, Chad; Witthuhn, Emily; Needham, Shane; Olson, James M
• Is Part Of: Bioanalysis. Volume 11:Number 6(2019)
• Page Start: 485
• Page End: 493
• Publication Date: 2019-03-19
• Subjects: μSPE -- cystine-dense peptides -- knottin peptide -- MFLC–MS/MS -- miniprotein -- MRM -- pharmacokinetics -- sample preparation -- therapeutic protein bioanalysis; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2018-0276
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: Develop a universal extraction and liquid chromatography-mass spectrometer method to simultaneously analyze cystine-dense peptide (CDP) miniproteins from rat and human plasma. The results of the analysis will be used to assist selection of therapeutic drug candidates from the vast CDP library.Methods & results: A micro-elution solid-phase extraction method was developed for the sample preparation of the CDP peptides in rat and human plasma followed by analysis by microflow liquid chromatography MS/MS. The methods developed for drug discovery were found to be accurate (±≤15.2% from nominal concentrations) and precise (≤13.4% CV), with a dynamic range of 1.00–500 ng/ml and extraction recoveries of 47.2–99.0%.Conclusion: This bioanalytical method can be utilized to screen CDP proteins and other miniproteins for drug discovery, candidate selection and further drug development.