The Importance of Tear Fluid Introduction

Tear is a complex mixture of proteins, lipids, mucins, water, and salts, performing multiple functions: i) Lubricate the ocular surface; ii) Provide nutrients to the corneal epithelium; iii) Provide a moist and smooth surface for healthy vision; iv) Protect the eyes from pathogens. Compared with other body fluids, tears are highly protein-concentrated and readily accessible, making tears a popular target for ophthalmology biomarker research. Tear fluid, as a source of potential biomarkers of ocular and systemic disease, may serve as a noninvasive strategy for detecting patients at increased risk or disease progression for eye or systemic disease compared with other bodily fluids, thereby helping improve diagnostic accuracy and understand disease pathophysiology.

Fig. 1. Tear biomarkers in dry eye disease. (Andreea Chiva, 2019)

Ace Therapeutics' Tear Fluid Biomarker Identification and Analysis Services

With the support of our advanced platform and experienced technical staff, Ace Therapeutics has developed a comprehensive tear fluid 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

• Tear Fluid Sampling

Ace Therapeutics' scientists use several established collection methods to obtain tear fluid, and selected the most appropriate sampling method according to the purpose of the research. Our tear fluid sampling methods include, but are not limited to.

• Cellulose Sponges
  A cellulose sponge is placed in the inferior conjunctival sac for approximately 1 minute to gather tear fluid. The method for collecting tear fluid has high efficacy and is non-irritating, as well as the ability to standardize the fluid collected.
• Schirmer's Strips
  Place the strip in the inferior conjunctival sac and leave in place until it is moistened to the control line. Subsequent incubation in a buffer solution rehydrates the bands so that proteins or metabolites can be extracted for further molecular testing. This technique yielded higher interleukin recovery compared to samples collected using cellulose sponges and improved protein identification compared to tear fluid collected using capillary tubes.
• Capillary Tube
  An end-to-end plastic capillary tube with a capacity of 10 μL is placed in the inferior conjunctival fornix to collect the desired volume of tear fluid by capillary action. Transfer the capillary to a perforated 0.5 mL tube and centrifuge into a 1.5 mL tube to recover tear fluid.

Note: Our scientists select the appropriate collection method for each specific study.

• Tear Fluid Proteomic Analysis

Our scientists use 1D and 2D gel electrophoresis (1DGE and 2DGE, respectively) for the separation, visualization, and size determination of proteins in tears, and MS for identification of protein-rich spots and relative quantification of proteins; Data-dependent acquisition (DDA) was used to identify peptides in unknown mixtures; Isobaric tagging for relative and absolute quantification (iTRAQ) and tandem mass tags (TMT) to quantify differentially expressed proteins in tear fluid. Thus discovering potential tear biomarkers.

• Tear Fluid Genomic Analysis

Our tear fluid genomic analysis employs automated workflows and platforms to generate reproducible results by providing genomic analysis-based PCR, qPCR, and Sanger sequencing for efficient and cost-effective genomic analysis. Our service overview is to extract RNA, DNA, and microRNA through sample preparation, and use PCR, qPCR, and NGS for tear gene sequencing analysis; Validation of gene expression levels in different ophthalmic disease models, and gene expression analysis and whole pathway analysis before and after drug treatment by targeted gene expression analysis; Predictive tear fluid biomarkers through gene fusion and gene mutation validation and qPCR-based SNP genotyping.

• Tear Fluid Epigenomics Analysis

Our tear fluid epigenomic analysis employs next-generation sequencing (ATAC-seq, DNase-seq, ChIP-seq, FAIRE-seq, MeDIP-seq, Small-RNA-seq), quantitative PCR for gene-specific analysis (methylation-specific, single miRNA, post-ChIP) and high-throughput screening to identify and elucidate the function and potential biomarkers of key epigenetic targets of aberrant growth signaling in ocular diseases.

• Tear Fluid Metabolomics Analysis

Tear fluid metabolites are often the end products of gene, mRNA and protein activity. Ace Therapeutics' scientists provide LC-MS based tear fluid non-targeted metabolomics analysis and targeted metabolomics analysis, and identify tear fluid biomarkers by constructing potential biomarker models. Our analysis process mainly includes sample extraction and purification, LC-MS detection, principal component analysis, metabolome enrichment and pathway analysis, and receiver operating characteristic (ROC) curves to identify potential biomarkers.

• Tear Fluid Lipidomic Analysis

After tear fluid samples are obtained, our scientists separate the analytes by liquid-liquid extraction or/and protein precipitation to reduce matrix effects, followed by reverse-phase high-resolution mass spectrometry (HPLC-MS) or/and direct injection mass spectrometry (DI-MS) for component analysis to discover and validate potential tear biomarkers.

Why Choose Us?

• Comprehensive multi-omic tear fluid biomarker analysis.
• Powerful biological information processing and data processing capabilities.
• Reliable data reproducibility, reducing batch effects.
• Research on disease mechanisms and drug mechanisms of action

At Ace Therapeutics, we continuously invest in cutting-edge technologies, validate and develop novel assays, and provide our clients with a comprehensive tear 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. Andreea Chiva. Tear Biomarkers in Dry Eye Disease. European Ophthalmic Review. 2019:13(1):21-6.
2. Zong Y, Cheng C, Li K, et al. Metabolomic Alterations in the Tear Fluids of Patients With Superior Limbic Keratoconjunctivitis. Front Med (Lausanne). 2022;18;8:797630.

• Conjunctival Biomarker Identification and Analysis Services
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