Analysis of Glycan Structure of Recombinant Protein Vaccines

Recombinant protein vaccines are a type of vaccines that do not contain complete pathogens and are prepared from specific protein antigens produced in heterologous expression systems. Recombinant protein vaccines have several advantages, including good safety, strong stability, and relatively low cost, and have been widely favored by researchers in recent years. In recombinant protein vaccines, glycan modification is a key parameter. This is because the structure of the glycan can affect the immunogenicity of the vaccine, thus affecting the efficacy of the vaccine. For example, certain glycan structures may increase the immunogenicity of the protein, while other glycan structures may reduce immunogenicity. The glycan modification may be influenced by many factors during the production process of the vaccine, including the type of host cell, culture conditions, protein folding processes, etc. Therefore, it is very important to perform detailed analysis and control of the vaccine's glycan modification.

 

Analyzing the glycan structure includes whether multiple different glycans are connected to specific sites, the order of arrangement of each sugar base, the conformation of the isoforms of the sugar base itself, the cyclic form of the sugar base, the connection method between the sugar bases, and the branching characteristics, etc. The analysis of oligosaccharide chains usually relies on the high-resolution mass spectrometry platform of liquid chromatography-mass spectrometry. First, PNGase F glycosidase is used to hydrolyze the glycan from the glycoprotein. Subsequently, the hydrolyzed glycans are derivatized and labeled with 2-AB reagent. After the labeling is completed, the glycan sample is injected into the liquid chromatography high-resolution mass spectrometry platform for data acquisition. The glycan analysis software is used to structurally analyze the collected mass spectrometry raw data. During the analysis process, the liquid chromatography equipment is equipped with a fluorescence detector, which can be used for quantification of the glycans. At the same time, the tandem high-resolution mass spectrometry system is used for structural analysis and identification of glycans. The liquid chromatography-tandem mass spectrometry method combines mass spectrometry and chromatography technology, which can effectively reveal the structure and characteristics of oligosaccharide chains.

 

MtoZ Biolabs provides drug quality research services that comply with global pharmaceutical regulations for customers. We use Thermo Fisher's Q ExactiveHF mass spectrometry platform combined with Nano-LC to provide a fast and efficient recombinant protein vaccine glycan analysis service package, which can qualitatively and quantitatively identify glycoproteins in mixed systems, including N-glycotypes, N-glycan structure analysis, and N-glycan content, etc., and can also provide customized analysis solutions according to demand to meet various testing needs. Feel free to consult.