Determination of glycosylation degree for glycoconjugate vaccines using a solid‐phase extraction combined with liquid chromatography and tandem mass spectrometry method

Abstract

In this study, a solid‐phase extraction with liquid chromatography and tandem mass spectrometry method was developed to determine the degree of glycosylation of glycosylation sites and the ratio of free carrier protein to total carrier protein for glycoconjugate vaccines. To remove and enrich the glycosylated peptides, a solid‐phase extraction method was developed, optimized, and hyphenated to liquid chromatography−tandem mass spectrometry. The developed solid‐phase extraction with liquid chromatography−tandem mass spectrometry method was shown to possess a wide linear dynamic range (0.03−100 μg/mL), a high sensitivity (0.03 μg/mL for CRM197), good interday and intra‐day precision (relative standard deviation of peak area < 3.3%), and good recoveries from vaccine matrix (90−105%). Finally, the method was utilized to determine the degree of glycosylation and free carrier protein to total carrier protein ratio for pneumococcal conjugate vaccines and meningococcal vaccines. For quality evaluation of glycoconjugate vaccines, the method could provide more information than the traditional size exclusion chromatography method. Fourteen and twelve reported glycosylation sites for CRM197‐ and tetanus toxin‐based vaccines can be detected, respectively.

Molecularly imprinted solid‐phase extraction of Chikusetsu saponin IVa from Panacis majoris Rhizoma

“…It combines separation and concentration using different solvents to selectively clean impurities or elute target substances with high purity. Among SPE systems with different kinds of sorbents, MIP-based SPE (MISPE) is a promising technique that provides excellent selective adsorption ability and applicability [19]. With separation specificity and high efficiency, MISPE has the advantages of simple operation, time-saving, labor-saving, and less environmental pollution, which can overcome the disadvantages of tedious pretreatment from a complex sample system [20][21][22].…”

Hydrophilic carrageenan functionalized magnetic carbon‐based framework linked by silane coupling agent for the enrichment of N‐glycopeptides from human saliva

“…Thanks to the high sensitivity and speed of detection, MS has become the most effective tool to accurately determine the glycosylation sites of proteins [11][12][13]. However, if MS is used to directly characterize proteins, some limitations will be exposed.…”

Simultaneous determination of capsid proteins in nine‐valent human papilloma virus vaccines by liquid chromatography tandem mass spectrometry

“…LC-MS/MS technology is widely used to quantify compounds in a complex matrix [8][9][10]. Previously, a LC-MS/MS method was developed for the determination of two types of L1s in a bivalent HPV-16/18 vaccine [11].…”