PEGylated protein can prolong the half-life of protein drugs and greatly extend the biological activity of protein drugs in organisms. The homogeneity of protein can ensure the accuracy of experimental results and accelerate the process of protein function research. Removal of contaminants and modified forms of protein can ensure the safety of protein drugs is a key link in the quality control of biopharmaceuticals.
Creative Proteomics is a biopharmaceutical research partner for R&D personnel with multiple experimental equipment platforms and biopharmaceutical research and development experience. We sincerely assist customers in accelerating the pace of biopharmaceutical research and development to accelerate the marketization process, and provide customers with high-quality proteomics-related analysis services. The ICH Q6B guidelines require protein purity and homogeneity analysis reports for biopharmaceuticals. We will provide you with PEGylated protein homogeneity analysis around the ICH guidelines and the US FDA issues 'Points to Consider' document.
We Can Provide but Not Limited to:
- Protein purity analysis
- Protein isoform analysis
- Protein hydrophobicity analysis
- Protein polymer analysis
- Protein fragment analysis
- Protein isoform analysis
- Protein molecular mass analysis
- Host cell protein analysis
Technology Platform of PEGylated Protein Homogeneity Analysis Service:
Creative Proteomics researchers can use hydrophobic chromatography (HIC), ion exchange chromatography (IEC), size exclusion chromatography (SEC) and spectrophotometry, LC-MS/MS Coupled with In-Source CID to effectively and accurately analyze the homogeneity of PEGylated proteins.
We provide the following analysis methods:
PEGylated protein homogeneity analysis
1) HIC
HIC can effectively separate the isomers of polyethylene glycol proteins, and is more suitable for some structurally identical structures that cannot be separated by ion exchange chromatography due to the same isoelectric point, but have different surface hydrophobicity. It can be divided into isomers, dimers and multimers, and the HIC can also be used for the analysis of polyethylene glycol protein degradation products to realize the homogeneity analysis of polyethylene glycol protein.
2) IEC
This method can realize the analysis of impurities such as components with the same molecular weight and sugar chain structural isomers in the PEGylated protein, and can realize high-resolution and rapid analysis of charge isomers.
3) SEC
SEC is the preferred method for detecting impurities with a molecular weight greater than that of PEGylated proteins. The combination of SEC and IEC analysis methods can more efficiently realize the homogeneity analysis of PEGylated proteins.
4) Spectrophotometry
This method uses the fluorescein method to determine the degree of polyethylene glycol modification. The product produced by the reaction between fluorescein and the amino terminal of the protein lysine residue has a specific light absorption, and the average is determined by measuring the fluorescence value excited by the protein mixture modification degree.
5) LC-MS/MS Coupled with In-Source CID
Pegylated proteins can undergo collision-induced dissociation in the ionization source (in-source CID) of the mass spectrometer, where peptide fragments produced by the source CID are used as replacement ions. The PEG concentration can be determined by the MRM of these replacement ions, and finally a quantitative analysis of PEGylated peptides can be obtained. The specific workflow is as follows:
Advantages of PEGylated Protein Homogeneity Analysis Service:
- Short time-consuming: The service has multiple first-class experimental platforms and mature analysis methods, which can quickly complete sample processing steps.
- High sensitivity: Mass spectrometry and other analytical techniques have the characteristics of high sensitivity and high efficiency, and can obtain high-quality analysis reports.
- High quality: This service provides complete and comprehensive analysis reports on sample purity and homogeneity, and guarantees the repeatability of the experiment.
- High accuracy: Mass spectrometry can obtain high-precision analysis information of polyethylene glycol sites.
- High-throughput: This service technology has a highly automated analysis platform that can analyze multiple samples of polyethylene glycol protein at the same time.
- Rapid turnaround time: 5-7 days to provide detail analysis report.
- Customized service: We can provide you with tailor-made services according to your needs, and you can also choose the analysis plan you need.
Creative Proteomics experts can provide customers with detailed PEGylated protein homogeneity analysis reports, including providing you with a comprehensive experimental design plan, related peptide map analysis, and the final protein homogeneity analysis report and other data reports. We sincerely provide you with proteomics analysis services to help you quickly complete your biopharmaceutical research and development projects.
References
- Gong J, Gu X, et al. Quantitative analysis of polyethylene glycol (PEG) and PEGylated proteins in animal tissues by LC-MS/MS coupled with in-source CID. Anal Chem. 2014.
- Lu X, Gough PC, et al. Elucidation of PEGylation site with a combined approach of in-source fragmentation and CID MS/MS. J Am Soc Mass Spectrom. 2010.
