Viral Glycoproteomics Services
It is well known that the surface proteins of most enveloped viruses are covered with glycans. In-depth characterization of glycosylation of viral glycoproteins has greatly promoted the understanding of viral pathogenesis as well as the development of vaccines and antivirals. As an international contract research organization (CRO) providing viral proteomic analysis services, Creative Proteomics is able to offer a series of viral glycoproteomics services, including N-linked gycoproteomics sample preparation, O-linked glycoproteomics sample preparation, and MS-based analysis of viral glycoproteomics. Our platforms are equipped with state-of-the-art facilities and highly experienced staff. We guarantee the finest results for our customers all over the world.
Glycosylation of viral glycoproteins
Enveloped viruses include some of the most common human pathogens that can cause infections of varying severity. Viral surface glycoproteins (VSPs) are tightly linked to the biogenesis of enveloped viruses, involving viral infectivity (virus entry, virus-host membrane fusion events), viral particle formation (assembly and release of progeny particles), and immune evasion. VSPs often hijack host cell's molecular machinery to obtain glycans. There are three main types of glycosylation, including N-linked, O-linked, and C-linked glycans, which are named after the acceptor amino acid atom to which the sugar moiety is linked. Among them, N-and O-linked glycosylation are the most common forms of glycosylation.
Fig1. Glycosylation patterns on enveloped viruses. (Bagdonaite, I.,et al, 2018)
Service Offering
Since glycosylation is macro- and micro-heterogeneous and changes the dissociation behavior of glycopeptides, specific sample preparation and tailored bioinformatic data analysis methods are required. Based on a team of professional and experienced scientists, Creative Proteomics is capable of offering N-linked glycoproteomic sample preparation services for mass spectrometric data acquisition.
Since the machinery of N-linked and O-linked glycosylation is different, specific types of sample preparation and analyses are required for each of these glycosylation types. We are proud to offer novel workflows for efficient O-glycopeptide generation for virus-infected cells. Our services mainly include the removal of N-linked glycans, the enrichment of O-linked glycopeptides, and the effective digestion of O-glycoproteins.
N-linked glycosylation is widely studied. The analysis of N-glycans in glycoproteins is an important branch of glycoproteomics. Based on a team of professional and experienced scientists, Creative Proteomics is capable of offering N-glycoproteomics services. Our services are based on advanced insturments, including MALDI-TOF MS and HILIC-UHPLC MS. Among them, MALDI-TOF is highly sensitive to the structure characterization of glycosylated compounds and can achieve high throughput detection. While HILIC is an emerging MS-compatible separation technique and can be applied to the analysis of N-glycan profiling.
We provide a powerful mass spectrometry (MS) -based strategy for globally mapping O-glycosylation sites on viral envelope proteins. Our services are based on a sequential enrichment with Vicia villosa lectin (VVA) and (peanut agglutinin) PNA coupled to tandem MS. The two-step lectin enrichment strategy is applied to capture desialylated O-glycopeptides in total proteolytic digests of virus-infected cells. Prior to performing the sequential enrichment, the major O-glycan structures produced during virus infection need to be identified. Moreover, we combine trypsin and chymotrypsin digestion to improve protein sequence coverage.
The advancements of MS-based proteomics have made it possible to perform a sensitive and comprehensive analysis of proteins and their post-translational modifications. So far, MS-based methods have been extensively used for the discovery and analysis of glycosylation on viral envelope glycoproteins. Based on the state-of-the-art facilities and highly experienced staff, we are able to provide information on glycan location, glycosite occupancy, and glycosite heterogeneities assignment for the study of glycosylation of viral glycoproteins.
Importantly, our viral glycoproteomics services include, but are not limited to the above services. If you are interested in our services, please don't hesitate to contact us. We are glad to cooperate with you!
Reference
- Bagdonaite, I., et al. (2018). “Viral glycoproteomes: technologies for characterization and outlook for vaccine design.” FEBS letters, 592(23), 3898-3920.
* For research use only.
