Vitreous Biomarkers and Retinal Disease

The vitreous humor, accounting for 80% of the volume of the eyeball, is a transparent gel-like tissue located between the lens and the retina at the back of the eye. The total volume is about 4 mL, 98% of which is water, and other components include collagen, hyaluronic acid, proteoglycan, and a small amount of vitreous cells. Among its functions, the vitreous contributes to the transparency of the intraocular medium, the maintenance of IOP, and the regulation of intraocular oxygen partial pressure. Furthermore, the vitreous provides protection by acting as a shock absorber, as collagen fibers reduce the compressive force of HA when the sphere is exposed to external pressure.

Numerous studies have shown the vitreous body is closely related to the pathogenesis of many vitreoretinal diseases, such as vitreoretinal traction which is the cause of macular hole, retinal detachment, proliferative vitreoretinopathy (PVR), proliferative diabetic retinopathy (PDR) and other diseases foundation. Recent scientific advances have allowed the study of many vitreous proteins as prognostic and predictive biomarkers for retinal diseases, including diabetic retinopathy (DR). ELISA, western blot and other methods are used to compare and analyze proteins in the vitreous under physiological and pathological conditions. They are widely used in the research of pathogenesis, early diagnosis, drug treatment targets and biomarkers of various diseases. In addition, the protein composition in the vitreous is relatively simple, and it is easier to detect subtle changes in the expression of low-abundance proteins, which makes it an ideal material for research into eye biomarkers in recent years. The study of vitreous biomarkers will help to further understand the pathogenesis of fundus diseases and provide an experimental basis for drug targets.

Fig. 1. Workflow of vitreous biomarker discovery by proteomics. (Komatsu H, et al., 2022)

Ace Therapeutics' Vitreous Biomarker Identification and Analysis Services

With the support of our advanced platform and experienced technical staff, Ace Therapeutics has developed a comprehensive vitreous biomarker analysis platform. Our ocular biomarker expert team has also perfected our technical pipeline for tear biomarker discovery. A variety of omics technologies are available for biomarker discovery, including proteomics, genomics, lipidomics, and metabolomics. In addition, a variety of high-throughput methods have also been applied, including mass spectrometry, protein microarray analysis, gene expression microarray analysis, and next-generation sequencing methods. Through our high-throughput system identification, functionally relevant tear biomarkers can be identified according to the needs of customers and used to support their clinical development.

Explore Our Service Strategies

• Vitreous Sampling

Obtaining a sample of vitreous humor is an invasive method. Our laboratory technicians obtain vitreous humor from research subjects (rats, mice, or other experimental animal models) through surgical vitrectomy. The collected vitreous samples were immediately stored in an ultra-low temperature freezer -80℃.

• Proteomic Analysis of Vitreous Biomarkers

Proteomic analysis of vitreous biomarkers is routine. The expert team at Ace Therapeutics pretreated the vitreous samples by digestion with a trypsin/Lys-C mixture and detected peptide fragments by liquid chromatography-mass spectrometry, and proteins were identified using a protein sequence database. Subsequently, with the help of powerful software in bioinformatics, various analyses including volcano plot, principal component analysis, cluster analysis, and functional interaction pathway analysis were performed to further identify candidate biomarkers. Finally, protein concentrations in the vitreous samples of the responder group were measured using ELISA to validate candidate protein biomarkers for the corresponding ocular disease.

• Analysis of Inflammatory Cytokine Biomarkers

Multiplex immunoassays are a powerful tool for analyzing low-volume body fluid samples. Several biomarkers associated with retinopathy have been found to be detectable in our previous studies, such as oxidative stress, hypoxia-inducible factors, angiogenic factors, pro-inflammatory cytokines, chemokines, and cell adhesion molecules. Ace Therapeutics provides our customers with comprehensive vitreous cytokine biomarker discovery and identification services through multiple immunoassay methods and other omics methods. This provides guidance for the research into corresponding retinal diseases.

• Metabolomic Analysis of Vitreous Biomarkers

Ace Therapeutics provides global customers with a variety of vitreous metabolism biomarker discovery and identification services. To obtain metabolic profiles and data, our scientists use liquid chromatography-mass spectrometry (LC-MS) to detect vitreous humor samples. By comparing the total ion chromatograms (TIC) and principal component analysis (PCA) results, and then compare whether there is a difference between the two groups of vitreous humor samples; then through T test analysis, cluster analysis, support vector machine discriminant analysis and other statistical analysis means to find important differential metabolites. Then, according to the mass-to-nucleus ratio and ion pattern of the metabolites, the structure of the metabolites was identified, and then the ROC curve was used to screen out the differential metabolites with strong discrimination ability as potential biomarkers, and explain their biological significance.

Why Choose Us?

• Analysis of multiple vitreous biomarkers.
• Powerful biological information processing and data processing capabilities.
• Reliable data reproducibility, reducing batch effects.
• Research on the mechanism of retinal diseases and the mechanism of drug action.

At Ace Therapeutics, we continuously invest in cutting-edge technologies, validate and develop novel assays, and provide our clients with a comprehensive vitreous biomarker service, relying on our flexible approach, extensive experience, well-established and validated workflows, highly dedicated bioinformatics team, extensive collection of historical data, high quality and fast turnaround to support your preclinical ophthalmic drug development. If you are interested in our services, please feel free to contact us.

Reference

1. Komatsu H, Usui Y, Tsubota K, et al. Comprehensive Proteomic Profiling of Vitreous Humor in Ocular Sarcoidosis Compared with Other Vitreoretinal Diseases. J Clin Med. 2022;11(13):3606.

• Tear Fluid Biomarker Identification and Analysis Services
• Conjunctival Biomarker Identification and Analysis Services
• Aqueous Humour Biomarker Identification and Analysis Services

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