The Importance of Aqueous Humor Biomarkers

Aqueous humor (AH), a clear slightly alkaline liquid formed and secreted by the ciliary body, occupies the space between the cornea and the lens. It is mainly composed of water (99.9%) with small amounts of carbohydrates, vitamins, proteins and other nutrients, as well as growth factors and cytokines. AH functions to support eye health by maintaining intraocular pressure, nourishing the vascular cornea and lens, and removing metabolic waste. The study of aqueous humor biomarkers helps to understand the mechanism of eye diseases and identify potential drug targets, while predicting the treatment response to eye diseases and increasing the chances of a clinical drug's success.

Fig. 1. Workflow of the human aqueous humor (AH) fluid proteome profile analysis and the discovery/validation of differently expressed proteins (DEPs) in the neovascular glaucoma (NVG) treatment. (Yu M, et al., 2020)

Aqueous Humor Biomarkers and Glaucoma

Studies have shown that the pathogenesis of glaucoma has not been clarified in clinical practice. The test of aqueous humor biomarkers can provide a basis for clinical research on the pathogenesis of glaucoma. For example, tissue inhibitors, matrix metalloproteinases, and transforming growth factor-β in aqueous humor can all be tested to determine trabecular meshwork damage. TGF-β is useful for analyzing cellular disease processes, including immune responses, wound healing, and cell proliferation. Glaucoma can be diagnosed and treated using aqueous humor biomarkers during the glaucoma research process. The accuracy of clinical diagnosis can be improved by analyzing the concentration of biomarkers in the aqueous humor. In addition, aqueous humor biomarkers are widely used in the evaluation of treatment effects, which can judge the prognosis of glaucoma surgery. Through the detection of PEDF and VEGF levels in the aqueous humor, the results of disease treatment can be effectively evaluated. It can judge and analyze the condition and treatment of patients with secondary glaucoma after vitrectomy and silicone oil tamponade for retinal detachment.

Fig. 2. Metabolomic biomarkers in the aqueous humor of POAG patients. (Tang Y, et al., 2021)

Ace Therapeutics' Aqueous Humor Biomarker Identification and Analysis Services

Ace Therapeuticss has a research team with decades of experience in ocular biomarker research, and we provide a variety of ocular biomarker development and validation services to global clients. Benefiting from our advanced analytical platform and experienced research team, Ace Therapeutics has developed a comprehensive analytical platform for the identification of aqueous humor biomarkers. We provide a one-stop service from the development of aqueous humor biomarkers in the early stage to the verification of aqueous humor biomarkers in the later stages. We can identify functionally relevant aqueous humor biomarkers according to the needs of customers and use them to support their clinical development.

As part of our service strategies, our scientists will utilize our multi-omics technology platform and high-throughput analysis methods to provide the highest quality services.

Explore Our Service Strategies

• Aqueous Humor Sampling

Unlike tear fluid, which is non-invasively obtained, aqueous humor sampling is usually performed surgically. During the study, for cases and control groups (in the case of rats), our scientists aspirated an appropriate amount of aqueous humor from the ocular stroma by corneal puncture, and aliquots of the aspirated AH samples were immediately stored in an ultra-low temperature freezer (−80 °C).

• Protein Biomarkers Analysis

Scientists at Ace Therapeutics provide customers with one-stop aqueous humor protein biomarker discovery and verification services, including protein extraction, protein digestion, LC-MS analysis, bioinformatics analysis, biomarker screening and biomarker verification. Throughout the service life, our expert team analyzes the differential proteins between responder and non-responder groups based on quantitative proteomics. Changes in protein abundance in a given group were first identified by employing a label-free quantitative analysis strategy of peptides, namely one-dimensional electrophoresis (1DE) and liquid chromatography (LC)-electrospray ionization (ESI)-MS/MS, followed by bioinformatics performing functional annotation and pathway analysis to determine the proteins of differentially abundant proteins, and finally analyzing and identifying differential proteins to determine potential biomarkers and validate them.

• Cytokine and Chemokine Biomarkers Analysis

Multiplex immunobead assays provide a versatile tool for the analysis of low volume body fluids. The application of antibodies provides high specificity, while Luminex microbeads offer the possibility of multiplexing, allowing the analysis of multiple analytes in one sample. Ace Therapeutics' scientists used multiplex immunobead assay technology for the analysis of cytokine and chemokine concentrations in aqueous humor samples. In light of this, we can screen the potential ocular biomarkers of aqueous humor and select the appropriate analytical method for verification based on the needs of the customer.

• Metabolite Biomarkers Analysis

Ace Therapeutics provides one-stop services from sample preparation, extract analysis, metabolite identification and quantification, and biomarker discovery. Our scientists use an LC-ESI-MS/MS system to analyze aqueous humor sample extracts and record metabolites. To assess the diagnostic power of certain metabolites (e.g., DEMs-A) as potential biomarkers. In the later stages of the analysis, our expert team will also conduct ROC analysis, using PLS regression, logistic regression, and random forest models analysis to judge the feasibility of DEMs-A as a potential biomarker.

• Oxidative Stress Biomarkers Analysis

The ocular surface is always exposed to strong light and solar ultraviolet radiation, and high metabolic activity will also increase the production of reactive oxygen species (ROS) and oxidative stress (OS). It has been hypothesized that the increase in antioxidant markers may reflect a local compensatory response of the eye to OS. Ace Therapeutics provides global customers with comprehensive services for the discovery and validation of aqueous humor oxidative stress biomarkers. Our scientists provide high-quality data by utilizing multiplex immunoassays with the help of next-generation sequencing (NGS) and genetic matrix analysis. We are able to detect TAC, MDA, ON, GPx and many other oxidative stress products, including but not limited to.

Why Choose Us?

• Analysis of multiple types of biomarkers in the aqueous humor.
• Powerful biological information processing and data processing capabilities.
• Reliable data reproducibility, reducing batch effects.
• Studies on the mechanism of glaucoma disease 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 aqueous humour 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.

References

1. Yu M, Xie F, Liu X, et al. Proteomic Study of Aqueous Humor and Its Application in the Treatment of Neovascular Glaucoma. Front Mol Biosci. 2020;7:587677.
2. Tang Y, Pan Y, Chen Y, et al. Metabolomic Profiling of Aqueous Humor and Plasma in Primary Open Angle Glaucoma Patients Points Towards Novel Diagnostic and Therapeutic Strategy. Front Pharmacol. 2021;12:621146.

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